WEBVTT

00:00.000 --> 00:09.560
Through where I think I'm at, now I'm going to give you my hypothesis in a nutshell and

00:09.560 --> 00:15.920
then I'm going to drink a beer and probably light a fire and see if I can read some comments

00:15.920 --> 00:18.520
before I go to bed.

00:18.520 --> 00:23.880
My current hypothesis is this, this likely originated from a laboratory source.

00:23.880 --> 00:29.000
Number one, laboratory leaks happen and you can look up many different articles including

00:29.000 --> 00:36.060
a few from Lynn Klotz who will show you definitively through history how leaks have happened through

00:36.060 --> 00:43.080
unintended infection, through waste products, lots of different ways and lots of different

00:43.080 --> 00:45.440
places it has happened.

00:45.440 --> 00:50.040
The second reason I believe this is true is because of the circumstantial bullseye, the

00:50.040 --> 00:56.160
idea that this broke out in Wuhan where the research on coronavirus was happening where

00:56.160 --> 01:01.360
the foremost groups were working on the foremost dangerous viruses and even though you want

01:01.360 --> 01:06.160
to say and people will tell you that this was BSL Level 4, most of the coronavirus work

01:06.160 --> 01:12.600
that is published in the literature is done in BSL Level 2 or BSL Level 3, even BSL Level

01:12.600 --> 01:19.960
3 is not a very strict biosafety level, I work in a BSL 3 level lab with my mice.

01:19.960 --> 01:24.600
And finally, the entangled financial interests I think are also very important to acknowledge

01:24.600 --> 01:28.280
here the amount of money that was given away to private equity, the amount of money

01:28.280 --> 01:35.040
that has been given away without context or identification to millions of corporations

01:35.040 --> 01:40.000
in America and the amount of money that is being given away through the Warp Speed Act

01:40.000 --> 01:47.400
that's allowing big pharma companies to make products that are guaranteed to be purchased

01:47.400 --> 01:52.560
or to be supported by government funding under the pretense that we need to bring as

01:52.600 --> 01:56.440
much of this to bear as possible.

01:56.440 --> 01:59.600
One of the things that you have to realize here, and I hadn't made this point yet but

01:59.600 --> 02:07.400
I know I'm glad I remembered, there is a federal law, the Defense Production Act, which allows

02:07.400 --> 02:14.240
the government to tell companies and corporations that they need to produce certain things at

02:14.240 --> 02:20.120
cost because it is for the benefit of all of the American people, for the benefit of

02:20.120 --> 02:22.280
America as a whole.

02:22.280 --> 02:26.560
This was an act that was designed to change the production of companies during World War

02:26.560 --> 02:34.200
One or World War Two and we could easily use this to direct pharmaceutical companies to

02:34.200 --> 02:40.800
develop specific things like, for example, produce hydrochloroquine or produce a vaccine

02:40.800 --> 02:46.880
of a particular design or a particular type and we could limit and control the amount

02:46.880 --> 02:52.800
of profit that was made so that nothing was, no one became rich from it.

02:52.800 --> 02:55.840
We could do the same thing with coronavirus testing.

02:55.840 --> 03:00.960
We could be using that act to make sure that no one is profiting from the testing, from

03:00.960 --> 03:06.400
the antibody testing and from the PCR testing, which I'm going to do another whole talk about

03:06.400 --> 03:12.640
that but PCR as a test is absurd right now and we know that because this test is not

03:12.640 --> 03:18.920
specific for SARS-CoV-2 but is only specific for SARS viruses in general and that means

03:18.920 --> 03:23.040
that we could, as I said before, be testing for the endemic version of the descendants

03:23.040 --> 03:27.680
of the original SARS, which are almost guaranteed to have gone through at least half of our

03:27.680 --> 03:32.120
population in the last decade and a half.

03:32.120 --> 03:36.200
So SARS-1 and its descendants have gone endemic, that's what I was just saying.

03:36.200 --> 03:42.560
The current PCR test is not specific for SARS-CoV-2 and that is known because it's the German

03:42.600 --> 03:46.480
company that originally produced it, knows that they've chosen a short sequence that

03:46.480 --> 03:47.480
they think is relevant.

03:47.480 --> 03:53.320
It is a short sequence that just comes from one of the open reading frame one proteins.

03:53.320 --> 03:58.520
We are being duped by a non-specific test and we would have gotten these numbers or

03:58.520 --> 04:03.880
numbers very similar to them with or without this pandemic because we've never looked for

04:03.880 --> 04:07.560
SARS viruses as they are spread through our population.

04:07.560 --> 04:09.600
We've just never looked.

04:09.600 --> 04:12.520
Nobody sampled for it until this year.

04:12.520 --> 04:19.000
It's a huge point to make because the SARS virus originally appeared in 2003 and disappeared

04:19.000 --> 04:25.120
in 2004 but it didn't really disappear, it just became not a problem.

04:25.120 --> 04:32.440
Descendants of that virus of various virulence and infectivity have circulated in the time

04:32.440 --> 04:40.440
since they didn't disappear and this test is not specific for any one of them and no one

04:40.440 --> 04:45.920
has ever done a study about how many descendants there are, how many people are infected,

04:45.920 --> 04:51.000
how many animals have been infected by it, etc.

04:51.000 --> 04:59.280
So that's a pretty interesting place to be in July of 2020.

04:59.280 --> 05:06.240
Thinking that there are descendants of the original SARS leak or leaks or zoonotic jumps

05:06.240 --> 05:12.240
which are building up an endemic signal in the background that could then be turned

05:12.240 --> 05:18.400
on in the flick of a switch, just kind of like Kevin McCurnan explained it in the video

05:18.400 --> 05:24.480
interview with me and a few other people where he said you could do it with HKU1.

05:24.480 --> 05:26.600
Just turn on the PCR and start monitoring it.

05:26.600 --> 05:32.360
You're going to see patterns of viruses moving through the population according to him.

05:32.360 --> 05:38.040
So if there are SARS descendants which could be used and misconstrued in that way, certainly

05:38.040 --> 05:41.520
that was a possibility that should have been considered from the very beginning of the

05:41.520 --> 05:44.520
pandemic and it was not.

05:44.520 --> 05:50.680
Certainly wasn't something that Kevin McCurnan brought up during his authorship of the paper

05:50.680 --> 05:57.560
that was let's say opposing the PCR test, opposing the PCR test by saying that some

05:57.560 --> 06:00.840
of the primers would make primer dimers.

06:00.840 --> 06:08.880
Never mind the whole pretense of being specific for a specific virus when the PCR test that

06:08.880 --> 06:15.520
was designed by the German lab was using sequences that they had found in Germany, sequences

06:15.520 --> 06:19.080
from German bats.

06:19.080 --> 06:26.280
I mean there was a lot of other reasons to lash out against the PCR test that was designed

06:26.280 --> 06:32.920
by Jorsten but primer dimers and this kind of thing was an odd sort of objection and

06:32.920 --> 06:38.080
if you look back and see who was on that paper, he will see that a number of early objectors

06:38.080 --> 06:41.440
were all roped in together.

06:41.440 --> 06:45.760
I think we're going to be coming back to that little gathering of quote unquote dissidents

06:45.760 --> 06:50.240
many many times in the next year or so.

06:50.240 --> 06:55.080
And so saying that we're being duped by a nonspecific PCR test is also really not specific

06:55.080 --> 07:01.080
enough even though right now this particular version of me was not completely queued in

07:01.080 --> 07:06.440
to the fact that this time that there could have been as many as 150 different variants

07:06.440 --> 07:14.640
of the test, 150 different variants of the test being utilized and being employed by maybe

07:14.640 --> 07:20.280
hundreds if not thousands of independent labs with various levels of quality control and

07:20.280 --> 07:23.120
certainly no oversight.

07:23.120 --> 07:27.520
And the crazy thing is as many of those small company labs that were doing a lot of the

07:27.520 --> 07:33.520
local testing especially in places where the testing was mandated, those are all gone.

07:33.520 --> 07:35.920
They don't exist anymore.

07:35.920 --> 07:39.800
Those companies took millions of dollars and they're gone.

07:39.800 --> 07:43.720
Their owners own houses and they're gone.

07:44.720 --> 07:50.360
And we're supposed to focus on the DNA fragments in the shot and forget about the fact that

07:50.360 --> 07:58.920
the whole the whole theater in the beginning to make worst case scenario real was wholly

07:58.920 --> 08:01.080
created out of out of fresh cloth.

08:01.080 --> 08:08.200
This is not sorry to start the show like this, but this is just the way it happened to be.

08:08.200 --> 08:12.880
I had this queued up for some reason.

08:12.880 --> 08:17.040
So the next thing that I want to say is our lack of biological knowledge and the general

08:17.040 --> 08:23.120
poor health of the of America, the American people is being used to create the crisis

08:23.120 --> 08:26.560
they need to divide and conquer us to ruin.

08:26.560 --> 08:27.560
Maybe America.

08:27.560 --> 08:28.560
I don't know.

08:28.560 --> 08:29.560
Crash the dollar.

08:29.560 --> 08:30.560
I don't know.

08:30.560 --> 08:34.480
Steel the rest of our are what limited treasury value we have left.

08:34.480 --> 08:36.720
I don't know.

08:36.720 --> 08:42.360
But I know for sure that they are combining our lack of biological knowledge and our general

08:42.360 --> 08:49.120
society's lack of good health and access to health care to create a crisis to usher

08:49.120 --> 08:54.640
in all kinds of changes that would otherwise never be necessary, and more importantly never

08:54.640 --> 08:56.840
be possible.

08:56.840 --> 09:01.200
Just like the Patriot acted 9-11.

09:01.200 --> 09:06.760
Highly profitable control mechanisms based on immunity are the near term goal.

09:06.760 --> 09:11.040
I believe this firmly now, there are too many people talking about it in the mainstream

09:11.040 --> 09:13.920
media hinting at it.

09:13.920 --> 09:18.600
Sound you hear is a single cicada, there are no crickets.

09:18.600 --> 09:23.000
Highly profitable control mechanisms, I'm talking about apps that you have to have and

09:23.000 --> 09:27.680
maybe even worse in order to cross state lines, in order to go to work, maybe in order to

09:27.680 --> 09:31.560
go to the store, you're going to have this happen.

09:31.560 --> 09:36.760
So there's going to be technology that needs to be distributed, there's going to be laws

09:36.760 --> 09:40.560
that are going to be need to be passed, there's going to be all kinds of products that need

09:40.560 --> 09:47.000
to be sold, and the whole idea is going to be to sell you on the idea that antibodies

09:47.000 --> 09:51.960
are the only evidence of immunity, and the only way to get antibodies is to get this

09:51.960 --> 09:57.840
vaccine, because a lot of people of course are not showing long lasting antibody levels,

09:57.840 --> 10:02.200
so even the people who've been infected, and that's what they're hyping right now is that

10:02.200 --> 10:06.760
you can get infected again, this is the narrative they're bringing to you so that they're going

10:06.760 --> 10:12.480
to tell you, the only way we're going to be sure is that if everybody gets vaccinated,

10:12.480 --> 10:20.360
and now we have billions of doses necessary, billions of doses backed by government money,

10:20.360 --> 10:28.320
backed by government mandate, controlled by apps, distributed yearly, we are talking

10:28.320 --> 10:33.560
about a very serious change in our healthcare system that will benefit the largest corporations

10:33.560 --> 10:38.400
and allow them to profit, because there are no controls on that profit right now, none

10:38.400 --> 10:42.560
at all, we have chosen to do nothing of the sort.

10:42.560 --> 10:47.320
That's why I'm sure this hypothesis is correct, the other reason why I'm sure this hypothesis

10:47.400 --> 10:51.920
is correct is, well I'm getting more sure that this hypothesis is correct, I don't want

10:51.920 --> 10:57.200
to sound like non-scientific here or non-objective, is because the guy that I first really got

10:57.200 --> 11:03.680
woken up by on the internet besides Dan of Harvard to the big house blog, is Wolfgang

11:03.680 --> 11:09.360
Wodach, and for some reason I never searched for this guy after I saw this German video

11:09.360 --> 11:14.520
that I had to get translated by a friend of mine, where he explained that it was likely

11:14.520 --> 11:19.000
a SARS virus or a SARS-descended virus that has been circulating viruses that have been

11:19.000 --> 11:28.000
circulating since the original SARS outbreak fizzled out in 2004 that we are testing for,

11:28.000 --> 11:34.840
and he only released that video in May or in April and it was only in German, and then

11:34.840 --> 11:35.840
I just never...

11:35.840 --> 11:40.320
I hope you see how significant it is that Wolfgang and I have now made contact in such

11:40.320 --> 11:42.360
a significant way.

11:42.360 --> 11:47.880
I hope you see what kind of forward progress is about to happen, don't be distracted by

11:47.880 --> 11:54.920
what's going on on Twitter and how people supposedly think that this story of the clones

11:54.920 --> 12:00.480
is an open and shut case now because there's effectively no swarm.

12:00.480 --> 12:06.300
He's been going around in circles on this idea for a very, very long time, and the reason

12:06.300 --> 12:11.240
why he's doing that is because all of RNA virology is dependent on it and no one can

12:11.240 --> 12:12.440
know that.

12:12.440 --> 12:21.840
If everybody understands that, that what they speak of as virology is a gross exaggeration

12:21.840 --> 12:28.800
of fidelity and it's a gross exaggeration of understanding, even the next strain database

12:28.800 --> 12:34.880
and phylogeny, even the Jed said database or whatever the hell it's called, all of these

12:34.880 --> 12:45.360
collections of genomes are presented to you as gold standard evidence of and proof

12:45.360 --> 12:49.360
of something for which they are not proof of.

12:49.360 --> 12:53.360
All we know for sure is that these people have managed to measure these sequences and

12:53.360 --> 12:58.580
a lot of these people believe these sequences are real, but we don't have any reason to

12:58.580 --> 13:04.360
believe that these sequences are somehow related to one another in a progression related to

13:04.360 --> 13:07.960
one another in an evolutionary way.

13:07.960 --> 13:13.280
We don't even know, no one is even talking about the possibility that these animals and

13:13.280 --> 13:20.600
ourselves would emit coronavirus like signals from our immune system or from the outside,

13:20.600 --> 13:24.320
from our lungs.

13:24.320 --> 13:30.600
No one's even talking about the possibility that these signals are being produced by the

13:30.600 --> 13:41.160
bacteria in our lungs or the bacteria in our airways, our GI tracks, wherever.

13:41.160 --> 13:46.600
No one's talking about the possibility that there's replication of RNA or DNA being carried

13:46.600 --> 13:51.400
by bacteriophages that's doing these signals, that's causing these signals, which really

13:51.400 --> 13:58.720
are just signals, genetic signals that are coincident with, well, genetic signals that

13:58.720 --> 14:03.240
are present and they're coincident with amplicons of PCR.

14:03.240 --> 14:11.920
It's very difficult at this size scale and at this sensitivity to definitively say that,

14:11.920 --> 14:18.520
well, this is definitely a something and we must always end up taking the word of somebody

14:18.520 --> 14:20.120
like Kevin McCurnan.

14:20.120 --> 14:21.280
They can't prove it to you.

14:21.280 --> 14:24.120
He's not going to ever teach you how this stuff works.

14:24.120 --> 14:27.760
He's always going to tell you that it's just too complicated for him to explain it

14:27.760 --> 14:31.280
to you, but I'm definitely wrong.

14:31.280 --> 14:38.120
When in reality, the idea that we're fighting about is really very simple and that's why

14:38.120 --> 14:43.600
it keeps spiraling out of control behind a block where we can't really have this discussion

14:43.600 --> 14:49.040
and why he keeps saying that we're not really, that he had this discussion a long time ago

14:49.040 --> 14:53.240
on email or he tried to do this on his sub stack before.

14:53.240 --> 14:57.160
I've addressed all the papers in his sub stack when they came out.

14:57.160 --> 15:02.120
I did a show about it several times in a row, which he completely ignored.

15:02.120 --> 15:03.120
And that's fine.

15:03.120 --> 15:04.120
He doesn't have to watch it.

15:04.120 --> 15:06.440
He calls my videos six hours Scooby-Doo videos.

15:06.440 --> 15:07.440
So that's fine.

15:07.440 --> 15:08.440
I get it.

15:08.440 --> 15:12.040
I should write a sub stack for him.

15:12.040 --> 15:13.880
We're going to figure this out, ladies and gentlemen.

15:13.880 --> 15:14.880
Don't worry.

15:14.880 --> 15:19.600
And when we figure it out the way that the ball will move forward, it will move in a very

15:19.600 --> 15:23.200
different way with a very different direction and a very different momentum.

15:23.200 --> 15:31.000
I assure you, I assure you, it's taken me a while to get this all straight in my head

15:31.000 --> 15:36.560
and make sure that we can move forward confidently.

15:36.560 --> 15:47.520
I've been trying very hard to remain true to my original idea, which was that this biology

15:47.520 --> 15:50.560
can't be impossible to learn.

15:50.560 --> 15:58.080
And any of the biology that underlies this phenomenon is worth learning.

15:58.080 --> 16:02.320
And so in the case of virology, I think all of us are just going to have to peel back

16:02.320 --> 16:11.040
our roll up our sleeves a little bit and learn some of this biology that they purport to

16:11.040 --> 16:16.600
have a monopoly on, that they purport to understand so well that you're just going to have to

16:16.600 --> 16:23.280
take their word for it, that they can't find any of this stuff in 2019 and before,

16:23.280 --> 16:26.280
but they sure can find a lot of it now.

16:26.280 --> 16:30.840
And that means that it's something going around and around and around and it couldn't

16:30.840 --> 16:34.060
be anything else.

16:34.060 --> 16:37.440
There are a few things in the background that we have to take and keep in mind.

16:37.440 --> 16:40.000
And there are a few things that we haven't addressed at all.

16:40.000 --> 16:45.680
And so it's just a real, it's a real mess right now.

16:45.680 --> 16:51.200
And so what they would like more than anything else for us to do is to rush as fast as we

16:51.200 --> 16:56.040
can thinking we know where we're going and thinking we know where the opening is to the

16:56.040 --> 16:57.040
cave.

16:57.040 --> 17:01.320
And we've got to keep remembering that we've got to keep our flashlights forward and we

17:01.320 --> 17:02.880
just got to keep working.

17:02.880 --> 17:11.560
It can't be distracted by the things that are going on, even if it is that you get fired

17:11.560 --> 17:26.800
or you get laid off or you get unexplainably your position is discontinued.

17:26.800 --> 17:34.280
And you are offered a non-disclosure agreement in exchange for $8,000.

17:34.280 --> 17:42.760
And you're left to kind of decide did I really put my neck on the line and put on

17:42.760 --> 17:55.240
the t-shirt and say that I was fighting for the truth and then now all of this hard work

17:55.240 --> 18:04.480
and sacrifice was worth a non-disclosure agreement and $8,000.

18:04.480 --> 18:08.640
I decided not to do that.

18:08.640 --> 18:13.760
And now I'm guessing I'm at the mercy of the internet for a little while.

18:13.760 --> 18:20.760
And we're going to try and make this work because I think actually this biology could

18:20.760 --> 18:23.600
save our kids.

18:23.600 --> 18:35.280
I actually think that if we can work through the literature and find truth in it, especially

18:35.280 --> 18:44.800
pre-COVID truth, and we can find biologists that we're working toward what they thought

18:44.800 --> 18:52.600
was the truth, I think we can find something that will be solid enough to stand on, that

18:52.600 --> 18:58.560
we can hand down to our children as an understanding of what these people lied about, what these

18:58.560 --> 19:02.080
people misled us about.

19:02.080 --> 19:08.000
And I think you're going to find that the illusion goes back farther than we thought.

19:08.000 --> 19:13.240
I think that's why so many of the interesting players in this are not like your typical

19:13.240 --> 19:21.400
players, like people who dropped out of grad school to start working on the Human Genome

19:21.400 --> 19:29.200
Project and then had like patents and companies and companies and ultra success.

19:29.200 --> 19:37.960
And then their boss went to the White House and was involved in the writing of $65 billion

19:37.960 --> 19:41.000
proposal for Warp Speed vaccines.

19:41.000 --> 19:49.920
And now he's been involved from the very beginning with objecting to the PCR test, but not PCR

19:49.920 --> 19:57.720
testing, and then all kinds of other random appearances for the RNA not being pure and

19:57.720 --> 20:06.920
then the RNA code on optimizing being responsible for different G quadriplexes and then came

20:06.920 --> 20:14.280
out later to be the double stranded DNA and the whole time this guy's been right there.

20:14.280 --> 20:19.920
Why has he been right there when his whole life is right now pot genetics and trying

20:19.920 --> 20:28.520
to, I don't know, I guess take over the commercial pot market by controlling genetics and virons

20:28.520 --> 20:37.640
and screening or providing testing materials for pot growers that will eventually become

20:37.640 --> 20:43.800
required by the government or some other scam.

20:43.800 --> 20:48.720
I'm sorry, but this is just this game has got to end.

20:48.720 --> 20:57.840
And these people, whether you like it or not, are not playing a small, tiny game.

20:57.840 --> 21:03.680
This is a game for all the marbles is for our grandchildren.

21:03.680 --> 21:08.720
That's why it seems so ridiculous.

21:08.720 --> 21:12.360
That's why it's being fought on Twitter.

21:12.360 --> 21:19.200
Do you think that Kevin McCurnan could have a real discussion about this and a group of

21:19.200 --> 21:23.960
people that all kind of knew about this stuff?

21:23.960 --> 21:28.960
Or would it would it really always have to be like him and Meryl Nast and then Andrew

21:28.960 --> 21:36.360
Kaufman and Christine Massey.

21:36.360 --> 21:42.800
The one time that we had a conversation a year ago, he couldn't really object to it.

21:42.800 --> 21:51.360
As you will see in tonight's recap, he just basically spit out exactly what I'm pointing

21:51.360 --> 21:55.640
out when everybody should be pointing out.

21:55.640 --> 22:04.240
That by making an infectious RNA and significant quantities, you could seed a coronavirus pandemic

22:04.240 --> 22:09.680
that was highly homogenous across many different places and maybe even would be highly stable

22:09.680 --> 22:11.440
for a little while.

22:11.440 --> 22:15.840
Yes, it's possible that clones transmit.

22:15.840 --> 22:20.760
I never said that they wouldn't.

22:20.760 --> 22:26.920
My main argument was that infectious clones and the whole principle of how they should

22:26.920 --> 22:33.520
or theoretically work is that you can create a much more homogenous viral stock that allows

22:33.520 --> 22:35.840
you to make a higher quantity of it.

22:35.840 --> 22:40.680
In fact, an infinite quantity of it is really only dependent on how to investment.

22:40.680 --> 22:45.760
If you wanted to do Jurassic Park, you could just do PCR construction of all the six or

22:45.760 --> 22:53.480
seven strands of DNA that you're going to use to ligate together and derive your RNA

22:53.480 --> 22:54.480
from.

22:54.480 --> 23:05.040
I mean, this guy's the limit if the idea is to just do it right.

23:05.040 --> 23:12.320
I'm just kind of, it's just kind of sad because this is where we're at, right?

23:12.320 --> 23:18.320
We have to protect our children from liars.

23:18.320 --> 23:24.800
Liars like Paul Offit, liars like Tony Fauci, liars like all of the people that work at

23:24.800 --> 23:31.200
the top levels of these pharmaceutical companies that know better.

23:31.200 --> 23:35.360
And we also have to protect our children from a bunch of people who don't know any better

23:35.360 --> 23:41.080
who aren't wise enough, smart enough, or I've turned the other way.

23:41.080 --> 23:44.280
In the military, they have caught what is called a lookaway doctrine.

23:44.280 --> 23:48.320
You know, if you're going to waterboard people, just let me close the door and get out of

23:48.320 --> 23:50.840
here before you start doing it.

23:50.840 --> 23:57.240
There are a lot of people involved in this little mystery that are also involved in that

23:57.240 --> 23:58.340
way.

23:58.340 --> 24:00.720
They know exactly how dangerous this was.

24:00.720 --> 24:04.980
They knew exactly how nasty this was going to get.

24:04.980 --> 24:08.800
And they knew that if they were going to get through this, they were going to break

24:08.800 --> 24:18.480
some eggs, but you know, make no mistake about it, ladies and gentlemen, breaking eggs

24:18.480 --> 24:26.960
is also going to require people on Twitter that have no business being on Twitter, talking

24:26.960 --> 24:32.400
about coronaviruses and fidelity of coronavirus replication and all this other stuff in their

24:32.400 --> 24:41.160
spare time while they're running companies on the genetics of weed.

24:41.160 --> 24:48.520
I mean, we are very, very, very close to exposing a group of people who some way or another

24:48.520 --> 24:53.840
were involved in the containing of the narrative at the same time they were involved in the

24:53.840 --> 25:01.680
perpetuating of the worst case scenario, making absolutely sure that the existence of the transmission

25:01.680 --> 25:07.400
of a novel virus was never questioned.

25:07.400 --> 25:15.200
And think about how strange it is that Eric Lander's former protege is the guy who was

25:15.200 --> 25:21.040
right there from the beginning, from the PCR objection to the RNA purity objection to the

25:21.040 --> 25:27.920
G quadriplex objections and the codon optimizing objections to the frame shifting objections

25:27.920 --> 25:32.920
all the way up to the double stranded DNA objections and what's really weird about the

25:32.920 --> 25:40.560
objections about the clones and about the proofreading in coronaviruses is that this

25:40.560 --> 25:46.360
thing that they recommended really, really early, that Cina Bavari predicted that they

25:46.360 --> 25:51.040
would use very, very early, that Rick Brighton knew that they were going to use very, very

25:51.040 --> 25:59.680
early, that Paul Cantrell knew that they were going to use very, very early, Remdesivir

25:59.680 --> 26:05.040
messes with the proofreading of coronavirus.

26:05.040 --> 26:08.240
So the proofreading of coronavirus is actually pretty significant.

26:08.240 --> 26:18.360
We need, we need to have high fidelity coronavirus replication that can be interfered with by

26:18.360 --> 26:25.840
Remdesivir and other antivirals so that the, the subtle balance between the mutagenesis

26:25.840 --> 26:32.400
in the quasi-species swarm and the stability of it is disrupted and you get this collapse

26:32.400 --> 26:35.240
of genetic stability.

26:35.240 --> 26:36.800
That's the story that they tell.

26:36.800 --> 26:38.720
That's the story that Mark Denison tells.

26:38.720 --> 26:41.800
That's the story that Cina Bavari told in his paper.

26:41.800 --> 26:44.200
That's the story that Ralph Barrick has told.

26:44.200 --> 26:50.760
And that's the story that is told when you argue that coronaviruses are very, very stable.

26:50.760 --> 26:56.000
You're arguing for Remdesivir's mechanism of action.

26:56.000 --> 26:57.000
Coincidence?

26:57.000 --> 26:59.000
Yeah, maybe.

26:59.000 --> 27:00.000
Believe what you want.

27:00.000 --> 27:01.000
What I want is high morbidity.

27:01.000 --> 27:02.000
I want people to complain.

27:02.000 --> 27:03.000
So what do I do?

27:03.000 --> 27:04.000
I go to Des Moines.

27:04.000 --> 27:05.000
Ladies and gentlemen, the people on the screen, I have nothing against Des Moines.

27:05.000 --> 27:06.000
I live there for four years.

27:06.000 --> 27:07.000
I go to Des Moines.

27:07.000 --> 27:08.000
I infect a couple of sentinel cases.

27:08.000 --> 27:09.000
I go to Seattle.

27:09.000 --> 27:12.000
I infect a couple of cases there.

27:12.000 --> 27:14.000
I go to North Carolina.

27:14.000 --> 27:15.000
I go to Wisconsin.

27:15.000 --> 27:23.000
What I'm doing is I'm using a dispersion methodology to be able to infect sentinel cases with a highly

27:23.000 --> 27:27.760
morbid condition.

27:27.760 --> 27:37.760
So I think what I want to do tonight is just quickly put up for a study hall.

27:37.760 --> 27:51.000
I've got this video queued up, a couple videos queued up on coronavirus replication, but

27:51.000 --> 27:56.640
I just want to look at...

27:56.640 --> 28:02.040
I just want to look at a couple things first to make sure that we still cover what we're

28:02.040 --> 28:03.040
really covering here.

28:03.040 --> 28:12.080
Let me escape out of this and start the new deck right away.

28:12.080 --> 28:19.200
I probably should just go really fast through it to bring everybody up to speed, but I hope

28:19.200 --> 28:20.800
you don't mind if I do that.

28:20.800 --> 28:28.640
So first of all, make sure you remember that one of the big, big, big things that is obvious

28:28.640 --> 28:35.240
to me as we move forward and we watch all these people scream and yell about how bad

28:35.240 --> 28:40.680
Jonathan Couey is for putting people's names on there or how bad it is that Jonathan Couey

28:40.680 --> 28:45.400
doesn't remember why somebody blocked him or whatever it is that they're crying about

28:45.400 --> 28:46.400
now.

28:46.400 --> 28:51.040
The one thing that you will remember and what I have been calling for more than a year now

28:51.040 --> 28:55.800
is that all these people will ignore this slide.

28:55.800 --> 28:59.960
You could be wrong about the clones, but right about this, and it wouldn't really matter

28:59.960 --> 29:01.360
now would it?

29:01.360 --> 29:03.840
And I assure you that I'm right about this.

29:03.840 --> 29:07.880
Intramuscular injection of any combination of substances with the intent of augmenting

29:07.880 --> 29:13.440
the immune system is dumb and transfection in healthy humans is criminally negligent.

29:13.440 --> 29:18.920
And by having me and you and all these people on Twitter that are trying to defend me,

29:18.920 --> 29:25.120
thank you very much, arguing about clones and whether or not the viral swarm is significantly

29:25.120 --> 29:30.200
diverse enough to support the idea that clones are different than the regular virus.

29:30.200 --> 29:34.440
This is all nonsense.

29:34.440 --> 29:38.400
This is all nonsense because this is true.

29:38.400 --> 29:43.560
This is all nonsense because this is true.

29:43.560 --> 29:49.120
And as long as they keep us occupied about whether or not J is right or wrong or whether

29:49.120 --> 29:54.720
it's okay to believe in J, then we're not going to we're not going to talk about this

29:54.720 --> 29:56.280
stuff.

29:56.280 --> 29:59.400
This is my most important message.

29:59.400 --> 30:04.880
This is the only message that I want to get to our children because if our children understand

30:04.880 --> 30:11.640
the big picture biology that underpins this statement, who cares about clones?

30:11.640 --> 30:17.040
Who cares how they do RNA virology and who cares if somebody with a Poch genetics company

30:17.040 --> 30:22.200
really does understand how they do coronavirus biology or not?

30:22.200 --> 30:31.680
Clearly he's being encouraged not to understand it because he is neglecting the very prominence

30:31.680 --> 30:40.440
of infectious clones from the early 90s on and the almost universal praise of how the

30:40.440 --> 30:45.640
development of infectious clones and their production has allowed the expansion of the

30:45.640 --> 30:51.120
understanding of how viruses work.

30:51.120 --> 30:56.400
We would have to be fools to believe that infectious clones weren't involved in this.

30:56.400 --> 31:02.400
They are the bedrock foundation methodology of all RNA virology, especially RNA virology

31:02.400 --> 31:08.520
around viruses that are difficult or impossible to culture for whom no infectious material

31:08.520 --> 31:12.800
is available.

31:12.800 --> 31:20.000
And so don't be fooled by somebody who can put up a few papers and then say a few sentences

31:20.000 --> 31:23.440
about why that paper makes them wrong.

31:23.440 --> 31:27.600
The time I've ever followed up on one of these papers, it doesn't really demonstrate

31:27.600 --> 31:36.520
what he says it demonstrates except for in a very sliver kind of way, but it's not addressing

31:36.520 --> 31:41.520
the very big pictures of what the natural phenomenon, what does the natural phenomenon

31:41.520 --> 31:45.520
look like is exactly the phenomenon that they don't want to talk about.

31:45.520 --> 31:50.400
They want you to believe and that's what he would like you to believe that the infectious

31:50.400 --> 31:59.360
clone replicates the natural phenomenon and that is a hundred percent wrong.

31:59.360 --> 32:04.920
And so they have fooled us into solving this Scooby-Doo and that's exactly what McCurnan

32:04.920 --> 32:06.440
is on right now.

32:06.440 --> 32:11.720
He wants you to believe that I'm supposed to have the exact explanation with all the details

32:11.720 --> 32:16.520
about exactly why the person came into my house, what they were doing there, what they intended

32:16.520 --> 32:20.280
to steal and why they were stealing it.

32:20.280 --> 32:24.840
And that's not the way a crime works.

32:24.840 --> 32:32.280
And our society, my family and yours are victims of a crime.

32:32.280 --> 32:36.120
We don't have any obligation as victims of a crime to know all the details and have a

32:36.120 --> 32:39.240
good explanation for everything.

32:39.240 --> 32:47.960
And we certainly aren't under any obligation to accept the explanation of the people who

32:47.960 --> 32:52.280
probably did it or are working with them.

32:52.280 --> 32:56.800
But we were fooled into believing that a worst-case scenario could have happened and we were fooled

32:56.800 --> 33:01.440
into believing that we could solve them, that we were watching people solve a mystery.

33:01.440 --> 33:08.880
That Rand Paul and Tony Fauci were arguing because they were on two sides of this mystery.

33:08.880 --> 33:12.560
And with this Scooby-Doo, the principle of informed consent has been ignored for the

33:12.560 --> 33:13.800
duration of the pandemic.

33:13.800 --> 33:22.560
Here we are trapped on our couch watching the whole thing unfold on TV.

33:22.560 --> 33:28.960
And so we've talked about how the pandemic was created from years ago up until now with

33:28.960 --> 33:35.520
a team of people or peoples around the world and especially in Western society that were

33:35.600 --> 33:39.600
there specifically to coordinate and amplify the worst-case scenario.

33:39.600 --> 33:47.440
The billions of people dead, the amyloid spike, the prion disease, the spike was killing cardiac

33:47.440 --> 33:50.840
cells, the whole nine yards.

33:50.840 --> 34:00.400
And the worst-case scenario was very cleverly tainted with aspects that they expected or

34:00.400 --> 34:04.120
feared would be a result of the transfection.

34:04.120 --> 34:07.280
They knew they were going to roll out the transfection so they knew that they could

34:07.280 --> 34:12.840
see this worst-case scenario with the worst-case scenarios of the transfection so that it would

34:12.840 --> 34:20.880
never really be clear to an unsuspecting and completely compliant populace that they were

34:20.880 --> 34:25.480
confounding the effects of the virus with the effects of the shot because you were given

34:25.480 --> 34:30.320
a viral protein and the viral protein turned out to be wicked.

34:30.320 --> 34:34.640
The people that were co-opted very early on were briefed that this was a national security

34:34.640 --> 34:40.360
thing and so you couldn't just talk about it.

34:40.360 --> 34:45.000
But if you go along with our little role here then maybe you can sell a book about Ivermectin

34:45.000 --> 34:52.760
or you can sell a book about hydroxychloroquine or you can sell a book about the lab leak.

34:52.760 --> 34:57.520
And in the end the toxicity of the virus, spike protein has been confounded with the

34:57.560 --> 35:03.520
toxicity of the transfection along with, of course, since we saw through this, since

35:03.520 --> 35:09.160
we saw through this little ruse a few years ago, a couple years ago, and we stopped believing

35:09.160 --> 35:12.760
this ruse.

35:12.760 --> 35:16.040
One of the reasons why we stopped believing this ruse was because we realized that the

35:16.040 --> 35:21.720
spike protein that's produced by a codon-optimized pseudo-uridine chemically altered RNA is not

35:21.720 --> 35:29.240
going to be anywhere structurally related in epitopes to that of one by the viral sequence.

35:29.240 --> 35:34.520
And we came to that conclusion after having Kevin McCurtain on our stream for two times.

35:34.520 --> 35:38.440
And so if the protein in three-dimensional structure is not equivalent to the protein

35:38.440 --> 35:43.040
that you're trying to build immunity to, chances are very good that the most damage

35:43.040 --> 35:48.040
associated immune epitopes there on that are also different.

35:48.040 --> 35:56.480
And so here we are, we know what the game was, we know how they did it.

35:56.480 --> 36:00.360
We know how they did it, we know they drove these mass casualty events and coordinated

36:00.360 --> 36:04.600
them in a way that made sure that nobody could question what actually happened.

36:04.600 --> 36:09.200
They co-opted a pre-selective group of narrative controllers and they probably brought more

36:09.200 --> 36:15.320
people on as the narrative went out of control as the compliance wasn't 100% with the uptake

36:15.400 --> 36:21.440
of the transfection, but a combination of background signal and nonspecific tests.

36:21.440 --> 36:24.640
And we don't have to figure that out for ourselves, we don't have to take everybody's

36:24.640 --> 36:27.160
word for it.

36:27.160 --> 36:32.720
And just because one person published a paper doesn't mean that that is the definitive

36:32.720 --> 36:39.720
Supreme Court evidence that now a scientific fact has been established, which is often

36:39.720 --> 36:43.520
how a lot of these rebuttal papers are presented.

36:43.960 --> 36:48.840
If you say somebody doesn't understand that the RNA, dependent RNA polymerase has a different

36:48.840 --> 36:55.320
replication capacity and then they cite a paper that doesn't immediately mean that

36:55.320 --> 37:00.920
that's the end of the discussion and that's really how this has gone from the very beginning.

37:00.920 --> 37:05.400
I think that they really thought that I was underprepared that I stumbled onto this idea

37:05.400 --> 37:08.960
and that's why the first three or four times that we had interaction with it, it was really

37:08.960 --> 37:11.080
pretty weak.

37:11.080 --> 37:16.280
And the one time we had an interaction live with my Kevin McCurnan and myself, I just

37:16.280 --> 37:21.440
kept kicking the ball back to him and at some point I just had to let him go because he

37:21.440 --> 37:27.040
just kept saying things that made it better.

37:27.040 --> 37:31.680
And so I think after that talk is when everything really peeled out of control and he decided

37:31.680 --> 37:36.520
that he had to make it appear as though he was slapping me down and used that stupid

37:36.520 --> 37:42.480
complicated strategy of his, you know, where he says, April back like six times and it

37:42.480 --> 37:47.360
forces everybody to Google it and so they all feel like fools and they don't want to admit

37:47.360 --> 37:52.920
that they don't understand what he's writing and then by not admitting that he's not teaching

37:52.920 --> 37:59.920
them anything, they let him get away with it, make it look like it's a consistent and consensus

37:59.920 --> 38:02.400
win on his part.

38:02.400 --> 38:08.880
And he refuses to acknowledge the fact that all of this stuff happened too.

38:08.880 --> 38:12.840
So there's actually really good reason to believe that there might not have been a real

38:12.840 --> 38:14.000
active spread.

38:14.000 --> 38:19.860
There's really good reason to be skeptical and that's part of what I find very funny

38:19.860 --> 38:24.720
about the one sub stack that actually dresses JJ's hypothesis in that sub stack.

38:24.720 --> 38:31.280
I think three times Kevin says maybe only twice that again, I'm not trying to decide

38:31.280 --> 38:36.040
here how much of the all-cause mortality should be attributed to COVID and how much

38:36.040 --> 38:42.120
of it should be attributed to bad ideas and man-demic.

38:42.120 --> 38:48.160
He makes the joke man-demic in his sub stack addressed to me from earlier this year, or

38:48.160 --> 38:55.720
maybe it's late last year, sorry, yes, it could be late 22 or early 23.

38:55.720 --> 39:02.800
And so I don't like to be accusatory, but in my perspective, we're dealing with someone

39:02.800 --> 39:05.080
who's behaving dishonestly.

39:05.080 --> 39:11.400
And from the very beginning, sort of a limited hangout kind of thing, giving as much, only

39:11.400 --> 39:16.560
as much information as to feel like he's giving a little bit, but never really bringing us

39:16.560 --> 39:17.840
all the way to the finish line.

39:17.840 --> 39:22.480
Are you telling me that Kevin McCurnan couldn't have imagined that there would be double-stranded

39:22.480 --> 39:26.800
contamination from the process two that he didn't know that they were going to use

39:26.800 --> 39:33.000
process to, that it was an option given the fact that Enovio was on BBC News with process

39:33.000 --> 39:36.600
two right behind them?

39:36.600 --> 39:38.480
I mean, how stupid do they think we are?

39:38.480 --> 39:42.160
How stupid does Robert Malone think we are that he doesn't know that process one and

39:42.160 --> 39:49.160
process two are very common differences that we knew that they made the joke on Twiv?

39:49.160 --> 39:55.240
Vincent Rancin-Yello talked about the amazing upgrade from nanograms to kilograms.

39:55.240 --> 39:59.880
I mean, come on, guys.

39:59.880 --> 40:05.840
And so these things all happened and they all want you to ignore the fact that they happened

40:05.840 --> 40:10.720
and they want you to continue to focus on a novel virus that's highly, high fidelity

40:10.720 --> 40:15.960
and that it can go around the world and change flavors repeatedly and with patterns.

40:15.960 --> 40:20.560
They want you to believe that passage of viruses gets you places.

40:20.560 --> 40:25.120
They want you to believe that when you stiff stuff together it gets you places.

40:25.120 --> 40:29.800
The worst-case scenario is that they made a lot of something and then they spread it

40:29.800 --> 40:30.800
around.

40:30.800 --> 40:33.880
That's the worst-case scenario.

40:33.880 --> 40:38.800
The worst-case scenario is that they made a lot of something that was pure and also toxic

40:38.800 --> 40:40.800
and fairly good at replicating.

40:40.800 --> 40:43.560
That would be awful.

40:43.560 --> 40:51.240
But it wouldn't be natural and none of the attributes of its spread would be natural

40:51.240 --> 40:55.000
because it would have started from a state that wasn't natural, a purity that wasn't

40:55.000 --> 41:01.440
natural and I can't understand why they insist on confounding this, ignoring this really

41:01.440 --> 41:03.640
easy thing.

41:03.640 --> 41:05.560
People asking me to write papers and stuff.

41:05.560 --> 41:10.160
It's like trying to write a paper about why cars use wheels.

41:10.160 --> 41:12.840
What does that mean?

41:12.840 --> 41:18.920
What does that mean when Alexandros Maranos and Kevin McCurnan say write up a paper?

41:18.920 --> 41:21.080
What does that mean?

41:21.080 --> 41:26.480
I'm telling you and it's a biological fact that the DNA can be copied with much higher

41:26.480 --> 41:33.280
fidelity than RNA and Kevin McCurnan even admitted it that you can make a lot of DNA

41:33.280 --> 41:36.720
and then you can make a lot of RNA from it and if you did that you would have a lot of

41:36.720 --> 41:37.720
genomic RNA.

41:37.720 --> 41:42.360
It'd be a little variable but it'd be a lot higher purity than anything you could ever

41:42.360 --> 41:49.400
achieve with any other method that we know and if you invested sufficient into the fidelity

41:49.400 --> 41:55.240
of it, use PCR methodologies, the state of the art technology, you could get purity levels

41:55.240 --> 41:58.240
heretofore on seed on earth.

41:58.240 --> 42:03.120
Distribute those everywhere and the signal would be brightest day.

42:03.120 --> 42:07.160
Not to mention the fact that you would have this all this leftover DNA that would also

42:07.160 --> 42:14.200
provide PCR signals because PCR is only looking for a very tiny amplicon so you'd

42:14.200 --> 42:20.920
have made tons of that amplicons too that you could distribute in sewers and in water

42:20.920 --> 42:25.280
and DNA is much more stable.

42:25.280 --> 42:30.840
That signal could potentially last for months and months depending on where you put it.

42:30.840 --> 42:35.640
Cruise ships could be positive for months if you had DNA that you distributed around.

42:35.640 --> 42:41.000
I mean come on ladies and gentlemen this is this is all obviously a joke thanks to

42:41.000 --> 42:44.120
James Giordano.

42:44.120 --> 42:49.080
Thanks to Kevin McCurnan, thanks to Kevin McCairn, thanks to Paul Catrell, thanks to

42:49.080 --> 42:56.920
Rick Bright, thanks to Tony Fauci, thanks to Ralph Barrick.

42:56.920 --> 43:01.060
You know one of the things that I think is really funny about Ralph Barrick is that actually

43:01.060 --> 43:07.700
I think he was a good guy and I think he's being co-opted.

43:07.700 --> 43:16.300
I want you to listen to this presentation that I think we are being set up to watch.

43:16.300 --> 43:18.460
That's all I'm going to say.

43:18.460 --> 43:24.300
I'm not sure if it's an authentic presentation, there are a couple times where it goes silent

43:24.300 --> 43:30.740
for like five or six seconds and I'm not sure it's an audio glitch, it could be an erasure

43:30.740 --> 43:35.020
and a couple times when it goes quiet they are talking about very key aspects of the

43:35.020 --> 43:37.660
infectious cycle.

43:37.660 --> 43:44.860
So this YouTube video was actually posted in a sub stack of Peter McCullough and this

43:44.860 --> 43:50.020
John O'leke or something like that and so that's how I found it and so I'm watching

43:50.020 --> 43:54.740
it for that reason but what I want you to remember is that this is in between SARS-1

43:55.740 --> 44:02.780
and Ralph Barrick may in theory, I want you to entertain this possibility, Ralph Barrick

44:02.780 --> 44:13.220
may still be an innocent bystander, a guy who's just part of the of coronavirus biology

44:13.220 --> 44:21.900
and he could still be in earnest thinking that coronaviruses could be one, a biotechnology

44:21.900 --> 44:32.780
to a pan vaccine technology and three, some kind of, I already said that, but some kind

44:32.780 --> 44:44.900
of future genetic modification technology so as they stand, some useful, then also modified

44:44.900 --> 44:50.980
and then also modified again I mean they were a huge potential in his mind and so I really

44:50.980 --> 44:56.380
think it's cool to listen to a video like this, even if it's altered, even if we're

44:56.380 --> 45:04.620
being tricked, I'm willing to bet that you're gonna hear enough of a guy who is fascinated

45:04.620 --> 45:12.500
by what he believes he is doing and I think he is doing it and that is studying synthetic

45:12.500 --> 45:14.180
viruses.

45:14.180 --> 45:19.020
So viruses that they can't culture, that they couldn't really get to grow in a lab and

45:19.020 --> 45:24.580
sustain themselves but has found another way to do it when we have these sequences

45:24.580 --> 45:29.780
and we decide on a consensus how can we do it and to really hear somebody explaining

45:29.780 --> 45:36.140
how they are developing this technology to assemble full coronavirus genomes, I'm confident

45:36.140 --> 45:43.260
it's gonna help us in terms of moving forward and really breaking the veil of this whole

45:43.260 --> 45:44.260
thing.

45:44.260 --> 45:53.380
Well, Carolina, you're gonna tell us about the synthetic genomics of SARS.

45:53.380 --> 45:59.820
So I also would like to thank the organizers for inviting me to talk, it's been wonderful.

45:59.820 --> 46:02.180
Now you are gonna be disappointed with the video here.

46:02.180 --> 46:05.380
Capture some new ideas that can be tried out on viruses.

46:05.380 --> 46:06.620
Well, this is not very loud.

46:07.620 --> 46:13.420
I'm gonna talk about today our basic short introduction into SARS biology.

46:13.420 --> 46:17.140
Talk about the synthetic resurrection and reconstruction of a variety of zoonotic SARS

46:17.140 --> 46:18.780
viruses and their applications.

46:18.780 --> 46:27.140
Okay, so already we're talking about synthetic reconstruction and he used the word resurrection.

46:27.140 --> 46:35.220
So resurrection would be from a sequence, from a partial sequence and it's very important

46:35.220 --> 46:40.740
to hear that because this is part of the reason why, again, clones are not just nothing.

46:40.740 --> 46:46.820
They're very special because the vast majority of coronaviruses cannot be easily cultured

46:46.820 --> 46:53.900
and when they are cultured, they only really barely culture it, they call it extremely

46:53.900 --> 46:59.660
low titer, but it's not really culturing that you can't make enough to share and it's

46:59.660 --> 47:00.660
not culturing.

47:00.660 --> 47:01.780
So they have a big problem.

47:01.780 --> 47:08.340
It's not just that they don't have, they don't have them in the right cell culture

47:08.340 --> 47:12.980
or it's not the right receptor, it's more complicated than that.

47:12.980 --> 47:19.580
And I think it comes down to the fact that infectious particles are oftentimes replication

47:19.580 --> 47:25.820
and competent and if you get at the heart of this, you're gonna find that that's really

47:25.820 --> 47:33.300
where this all peters out, where the rubber meets the road is right there and that's

47:33.300 --> 47:35.620
where they just do the hand wavy.

47:35.620 --> 47:38.620
And that's why the no virus people had so much to stand on.

47:38.620 --> 47:46.460
That's why I think they were so dangerous and I think that's why they were so confusing.

47:46.460 --> 47:52.020
But are we, do you feel the pendulum swinging back now, do you feel it?

47:52.020 --> 47:56.740
Where we were almost all the way to no virus and really exploring the possibility that

47:56.740 --> 48:00.820
wow, did they really only put clones in like four people?

48:00.820 --> 48:05.300
Like Sohomish County man and a couple other people and sequence those and then the rest

48:05.300 --> 48:09.380
is all nonsense, it's possible.

48:09.380 --> 48:16.420
But there's a whole possibility space that expands from that very tiny number of people

48:16.420 --> 48:23.540
that were infected with clones that never went anywhere to another medium possibility,

48:23.540 --> 48:29.140
which is that they actually did put a significant a number of and concentration of clones in

48:29.140 --> 48:34.340
several places so that several people or many hundreds of people would be infected with

48:34.340 --> 48:37.940
a very pure infectious RNA.

48:37.940 --> 48:42.660
And I think over the next few days, as we explore these papers together, you're going

48:42.660 --> 48:49.940
to find a preponderance of evidence as I have, which indicate that these RNAs all by themselves

48:49.940 --> 48:53.220
do some pretty cool stuff.

48:53.220 --> 49:05.460
And as far as most virology papers are concerned, these are better than or more reliable than viruses.

49:05.460 --> 49:09.860
And what they do and how they do it is more reliable and you're going to figure out that

49:09.860 --> 49:18.500
that's because of the purity of the RNA, the purity of genomic RNA that gets into each cell

49:18.500 --> 49:27.540
is several orders of magnitude higher than the amount of genomic DNA that gets into a cell

49:28.180 --> 49:30.740
during passage.

49:30.740 --> 49:35.780
And that's what's really extraordinary here. That's really what's extraordinary here.

49:36.420 --> 49:40.420
And therapeutic and vaccine design.

49:40.420 --> 49:45.220
I'll also touch on codons, the optimization as a way to attenuate SARS pathogenesis,

49:45.220 --> 49:49.940
but I certainly haven't gone to the depth of studies that Eckerd's group has done.

49:49.940 --> 49:55.620
And then I'll talk about rewiring SARS-Coronavirus transcription circuits as a way universal strategy

49:55.620 --> 49:59.380
to attenuate viral pathogenesis.

49:59.380 --> 50:03.060
So this is a schematic diagram of the SARS-Coronavirus particle.

50:04.020 --> 50:06.420
It's about 100 nanometers from diameter.

50:06.420 --> 50:09.940
It contains a helical nucleic acid inside.

50:09.940 --> 50:12.820
It contains a single-stranded plus polarity RNA genome.

50:14.020 --> 50:17.140
This nucleic acid structure is surrounded by a lipid bilayer.

50:17.780 --> 50:20.500
It contains several viral glycoprotein spikes.

50:20.500 --> 50:25.060
The important ones for today's talk include the M glycoprotein and the E protein,

50:25.060 --> 50:26.260
which are shown right here.

50:26.820 --> 50:30.900
These proteins are absolutely essential for maturation and release

50:30.900 --> 50:32.580
and the production of virus particles.

50:33.220 --> 50:34.260
So keep that in mind.

50:35.220 --> 50:39.540
The second viral protein of interest for today's talk is the escolycoprotein gene

50:39.540 --> 50:43.460
that's shown right here, which gives the virus its unique occurrence in the electron microscope.

50:44.740 --> 50:48.820
It's the viral passion protein that binds to the receptor to mediate,

50:48.820 --> 50:50.180
docking, and entry into the cell.

50:51.140 --> 50:55.460
It regulates tissue tropism, species specificity.

50:55.460 --> 50:58.340
It contains a large number of important neutralizing epitopes,

50:58.340 --> 51:00.900
and it's the principal component of protective immunity.

51:01.860 --> 51:08.340
Now, if you tear this virus particle apart and look at the viral genomic positive-stranded RNA,

51:08.340 --> 51:11.060
it's about 30,000 base pairs in length.

51:11.060 --> 51:14.180
It's the largest plus polarity RNA genomes in nature.

51:14.180 --> 51:17.940
The first 20,000 base pairs are shown to encode the replicase proteins.

51:17.940 --> 51:22.340
So the job it is to replicate the genome, replicate the genome,

51:22.340 --> 51:26.740
and express sub-genomic messenger RNA that encode these downstream open reading frames.

51:31.540 --> 51:33.540
Here's the first blank space.

51:33.540 --> 51:37.460
Now, these downstream morphs encode the structural genes like the SEM and

51:37.460 --> 51:40.980
nucleocapsid protein that I just mentioned that were present in the virion

51:40.980 --> 51:43.220
in a variety of accessory morphs.

51:43.220 --> 51:48.020
To get these expressed in cells, you find a nested set of sub-genomic messenger RNA.

51:48.580 --> 51:52.580
They're ranged from the three prime end of the genome so that all the sequences in the smallest

51:52.580 --> 51:56.260
message are in the next largest message and so on and so forth.

51:56.260 --> 52:01.620
This organization allows different open reading frames to be placed at the five prime ends of

52:01.620 --> 52:06.100
different messenger RNAs so they can be efficiently translated and expressed in cells.

52:06.820 --> 52:07.700
Now, the trans...

52:09.460 --> 52:11.140
Here, again, is another dropout.

52:13.540 --> 52:13.940
I don't know.

52:13.940 --> 52:16.740
How 70 nucleotides shown here are these little blue boxes.

52:18.260 --> 52:21.460
That is derived from the five prime end of the genome.

52:21.460 --> 52:25.620
So these are discontinuous stretches of RNA that are joined going transcription.

52:25.700 --> 52:29.940
And these little red box elements right here are absolutely essential

52:29.940 --> 52:32.420
for mediating the joining of these two sequences.

52:33.380 --> 52:36.020
This will become very important later on in the talk.

52:36.020 --> 52:36.980
So keep the...

52:36.980 --> 52:41.940
Okay, so what he's explaining here just to kind of to bring you up to speak.

52:41.940 --> 52:44.020
You can't really see it here and I apologize.

52:44.020 --> 52:46.740
I told you it was going to be a bad... the graphics are bad.

52:48.660 --> 52:51.060
It's weird. This was uploaded last year.

52:51.060 --> 52:52.740
I can't find another copy of it.

52:53.620 --> 52:58.820
There's those two things that were edited out there or blanked out in the beginning there.

52:58.820 --> 53:02.100
I can't tell but it sure feels like there was an erasure there.

53:02.660 --> 53:07.700
And he's talking about these TRSs which are translation regulatory sequences.

53:08.660 --> 53:12.660
They are small sequences that are repeated throughout the genome.

53:13.380 --> 53:17.140
And it's not very clear to me upon reading the genome...

53:17.780 --> 53:24.660
or sorry, the literature exactly how these transcriptional regulatory elements

53:25.380 --> 53:26.660
interact with one another.

53:26.660 --> 53:36.900
But it feels like, if I can give you my best impression, it feels like that these TRS segments

53:37.620 --> 53:44.740
are repeated because the RNA molecule has a three-dimensional interaction with itself

53:45.300 --> 53:50.260
and makes it not so that this part gets looped over to this part.

53:50.820 --> 53:56.580
And so then the RNA-dependent RNA polymerase can make a very short RNA that goes bloke

53:56.580 --> 53:57.860
and then it goes right to here.

53:58.740 --> 54:01.540
And it goes bloke and then it goes right to here.

54:01.540 --> 54:06.020
And so every time it gets to a translational regulatory element,

54:06.660 --> 54:12.420
there's the potential for this part of the five prime end to be looped in over here.

54:12.420 --> 54:16.020
And so then the RNA-dependent RNA polymerase that's translating it can

54:16.980 --> 54:19.300
to make this sub-genomic RNA.

54:19.300 --> 54:21.620
And that's probably completely wrong.

54:21.620 --> 54:26.180
But I do know that the translation regulatory elements are all the same.

54:27.140 --> 54:33.940
And it is from these spots in the genome that the recombination is most likely to occur.

54:35.780 --> 54:40.020
And one of the ways, and Ralph Berwick will explain it in this talk later,

54:40.100 --> 54:44.420
that they thought to make an attenuated virus that could not

54:45.220 --> 54:49.860
de-attenuate by recombining with other viruses in the wild

54:50.660 --> 54:56.980
was to artificially change the translation regulatory segments of the artificial genome

54:57.940 --> 55:04.980
so that this virus, when co-infecting a cell, would have translation regulatory elements that

55:04.980 --> 55:09.700
were incompatible with the genomes that would be present in the wild.

55:09.700 --> 55:17.620
And therefore they could not rearrange their genome to acquire genes from wild coronaviruses.

55:17.620 --> 55:23.460
So what he's explaining here is an aspect of the coronavirus genome that if you take their

55:23.460 --> 55:29.700
word for it, as I'm doing right now, is part of the regulatory mechanism by which the

55:30.660 --> 55:36.500
entire signal genome actually gets translated into these sub-genomic RNAs that we've been talking

55:36.500 --> 55:41.540
a lot about and that we understand are in abundance of several orders of magnitude more

55:41.540 --> 55:47.300
abundance than the full genome, whenever they try to find the total RNA in any of these preparations.

55:47.300 --> 55:53.940
I hope that helped a little bit, especially because you can't see Jack in this sort of blurry thing.

55:53.940 --> 55:58.180
But we are listening to the great RB, so it's pretty fun.

55:58.740 --> 56:04.020
The concept of TRS elements, a little red box element, is being essential for regulation of

56:04.020 --> 56:12.420
messenger RNA synthesis. Now, let's talk about synthetic genomics in the context of a platform

56:12.420 --> 56:17.780
to control emerging infectious diseases. If you think about emerging viruses, they have tremendous

56:17.780 --> 56:23.940
potential to cause high morbidity and mortality. They have tremendous potential to cause high

56:24.020 --> 56:31.220
morbidity and high mortality, meaning that there are small examples of this happening. But again,

56:31.220 --> 56:38.100
it's the pandemic potential is different. I'm sure that if you had enough clone of a clone

56:38.100 --> 56:42.660
of an RNA like this and you put it in somebody's lungs, yeah, or squirted it in some,

56:42.660 --> 56:49.940
an animal's nasal passages, you'd have some problems. If you put that RNA in a cell culture and then

56:49.940 --> 56:54.580
passage it a few times, you're going to have a lot of RNA in every cell. It's all going to be

56:54.580 --> 56:58.820
the same. It's all going to do its viral thing. It's all going to package a bunch of stuff up.

56:58.820 --> 57:05.780
You're going to have a huge concentration of highly homogenous RNA particles, as far as we can tell,

57:05.780 --> 57:12.500
viral particles, exosomes, whatever, that you would not have if you just started with a sample from

57:12.500 --> 57:17.300
the bat and then started passageing it and tried to make a lot. And that's the point.

57:21.220 --> 57:26.900
Let's let it keep playing here. Maybe I should just quick, just let me give you this one thing here.

57:26.900 --> 57:31.220
I had this queued up and then I just, I just realized we were going way, I was going way off

57:31.220 --> 57:43.060
balance here. But here's a mini review from a virology in 1994, infectious transcripts,

57:43.060 --> 57:50.020
CDNA clones of RNA viruses. Recombinant DNA technology makes it possible to analyze and modify

57:50.020 --> 57:55.540
genomes at the molecular level and thus gain deeper insight into their organization and expression.

57:55.700 --> 58:01.140
Oops, sorry. In this, in this respect, viruses because of the small size of their genome are

58:01.140 --> 58:06.820
particularly amenable to such investigations. In spite of this, the study of molecular biology of

58:06.820 --> 58:13.860
non-retroviral RNA viruses has long been hampered by the fact that these viruses do not encompass a

58:13.860 --> 58:20.740
DNA intermediate step in their replication cycle. Therefore, since to date, the extremely varied

58:20.740 --> 58:26.020
and powerful molecular biology techniques aimed at modifying nucleic acids have been directed

58:26.020 --> 58:31.780
essentially at DNA substrate, new molecular tools had to be developed. The possibility of

58:31.780 --> 58:42.260
obtaining infectious clones as CDNAs or as in vitro transcribed RNA copies, which is what an RNA

58:42.260 --> 58:50.900
infectious clone is. Corresponding to the genomes of RNA viruses has greatly enhanced the

58:50.900 --> 58:56.100
potential of investigations. Indeed, they can facilitate studies of viruses that are present

58:56.100 --> 59:07.940
only in low titers in infected cells or whose isolation is problematic. Interesting. That's from

59:08.340 --> 59:20.820
1994. Let's go to 2019 from Cina Bavari and Alison Totura, a former postdoc of Ralph Barrick.

59:22.340 --> 59:30.100
Section 2, 2.1, reverse genetic systems. Advances in the study of highly pathogenic coronaviruses and

59:30.100 --> 59:35.460
potential pan-coronavirus drug tech candidates partially depends on the technology to genetically

59:35.460 --> 59:40.180
manipulate coronaviruses to probe mechanisms of viral pathogenesis and antiviral drug activity.

59:40.820 --> 59:46.740
Reverse genetic systems synthetically generate viruses from known viral sequences. In situations

59:46.740 --> 59:52.500
who wear clinical isolates of infectious material unavailable and due to restrictions for collecting

59:52.500 --> 59:57.140
patient samples, shipping infectious materials, availability of containment laboratories,

59:57.140 --> 01:00:03.220
reverse genetic systems, provide the essential research materials for studies on viral pathogenesis

01:00:03.300 --> 01:00:09.860
in model development. Prior to the SARS pandemic, robust reverse genetic systems to manipulate the

01:00:09.860 --> 01:00:17.140
genomes of Scarsco V2s, sorry, of coronaviruses had already been developed by systematic assembly

01:00:17.140 --> 01:00:23.060
of cDNA consents to full-length infectious clones, allowing precise and genetic targeted

01:00:23.060 --> 01:00:29.220
manipulation of viral genes. More importantly, to me, infectious clones allow the creation of

01:00:29.220 --> 01:00:36.900
near homogenous viral stocks where the traditional viral stocks are prepared by

01:00:36.900 --> 01:00:44.660
amplification of infectious material in cell culture. Infectious materials, that's the stuff

01:00:44.660 --> 01:00:51.460
they take from bats, that's the stuff they take from sick people. Over many passages, strategies

01:00:51.460 --> 01:00:56.580
to build reverse genetic systems were rapidly applied to both SARS and MERS within the first

01:00:56.580 --> 01:01:03.700
year of identifying these viruses. In addition to reconstructing epidemic,

01:01:03.700 --> 01:01:09.780
epidemic strains of SARS-CoV-2 with reverse genetic systems allow targeting of mutations

01:01:09.780 --> 01:01:15.860
to specific viral genes and assembly of these virus when infectious material is not available.

01:01:15.860 --> 01:01:28.260
So they've seen pretty significant for RNA virology in general, but for some reason or another, a guy who

01:01:30.980 --> 01:01:35.220
was at the White House when he was what, 25 or something like that? I don't know,

01:01:35.220 --> 01:01:40.420
how old was he when he was at the White House for the first time and probably because of his dad,

01:01:40.500 --> 01:01:49.300
maybe, I don't know, but and then his, he drops out of grad school, goes to work for Eric Lander,

01:01:50.260 --> 01:02:01.300
Eric Lander's in the Biden White House, running stuff for the mRNA, and this guy's involved from

01:02:01.300 --> 01:02:06.660
the very beginning from the PCR shot all the way through all the objections that were ever made

01:02:06.660 --> 01:02:17.220
about the transfection and titrating them one by one and we're supposed to believe that

01:02:17.220 --> 01:02:21.860
clones are just like everything else. It's no big deal. Clones don't do anything. Clones are

01:02:21.860 --> 01:02:31.780
chemtrails stupid. It's pretty phenomenal, really, if you think about it. And severe economic

01:02:31.780 --> 01:02:37.780
hardship, good examples are HIV, the 1918 flu, far as coronavirus, H5N1,

01:02:37.780 --> 01:02:43.220
chicken and guinea virus, probably most recently. Now it's clear that zoonotic introductions are

01:02:43.220 --> 01:02:50.260
increasing and a large number of these viruses have large reservoir pools of animal strains

01:02:50.260 --> 01:02:56.420
like SARS and H5 that are maintained as sort of heterogeneous swarms of pools of viruses

01:02:56.420 --> 01:03:01.220
of which someone may actually emerge in the future to cause epidemic disease in humans.

01:03:01.860 --> 01:03:07.140
This raises a conundrum in terms of how do you protect the public health against the future

01:03:07.140 --> 01:03:14.340
occurrence that is hard to predict. I wonder how we would differentiate between a pathogen in bats

01:03:14.340 --> 01:03:21.460
that doesn't make bats sick. How would we differentiate that from a immune signal?

01:03:22.900 --> 01:03:31.380
Like let's say that the bats physiology is to emit coronaviruses. Let's say that a lot of

01:03:31.860 --> 01:03:44.180
mammals emit self replicating RNA packaged in a lipoprotein coat. Just just imagine that for a

01:03:44.180 --> 01:03:51.300
second. Is that crazy? Is that a crazy idea? Are we crazy? Are we a little crazy to assume

01:03:52.100 --> 01:03:57.780
that these RNA molecules coated in a lipid dental, a lipid protein, a lipoprotein coat,

01:03:58.020 --> 01:04:03.060
aren't we a little? Isn't it a little weird to assume that these guys are just all independent

01:04:03.060 --> 01:04:12.740
actors that are out of the control of the bats, out of the influence of the bats,

01:04:12.740 --> 01:04:20.660
out of the hands of the, despite the bats best effort, these coronaviruses are thriving inside

01:04:20.660 --> 01:04:23.620
of the bat population. Why does it have to be like that?

01:04:28.260 --> 01:04:31.300
Is it, is it really like that? Are we sure it's like that?

01:04:34.340 --> 01:04:41.460
Are we sure that Simeon virus 40 was really a virus that the monkeys body really wanted to get

01:04:41.460 --> 01:04:47.300
rid of but couldn't get rid of? Are we assuming that the pangolans have coronaviruses that they

01:04:47.300 --> 01:04:54.020
don't want that they're not supposed to have but they're just there because despite their best

01:04:54.020 --> 01:05:05.060
efforts, that's a huge assumption, isn't it? As opposed to it being like a signal or an offgassing

01:05:05.060 --> 01:05:10.660
or a noise, I like a signal. I like the idea of it being a signal.

01:05:11.460 --> 01:05:15.940
I don't like the idea so much of it being a pathogen anymore.

01:05:17.940 --> 01:05:22.900
But I definitely like the idea that Ralph Barrick thinks it's a signal and thinks that what he's

01:05:22.900 --> 01:05:27.620
working on is important and what he's working on is something that's worth understanding and that

01:05:27.620 --> 01:05:33.540
he's made a major breakthrough by figuring out how to make these full genomes and start there.

01:05:34.340 --> 01:05:40.820
I think it's possible. I think your your sliver of understanding, your sliver of specialty could

01:05:40.820 --> 01:05:47.060
be so great that you could get so fired up about making machine guns that you just don't realize

01:05:47.060 --> 01:05:55.460
that, wow, I made really, I mean this machine gun is pretty amazing and I'm proud of all the the

01:05:56.340 --> 01:06:02.100
the ingenious ideas I put into it but I'm a little disappointed with what it did to them all.

01:06:03.060 --> 01:06:05.700
That's possible. It's totally possible.

01:06:08.420 --> 01:06:13.220
You know, I was a clock maker but then I got into guns and I just figured out this thing and it

01:06:13.220 --> 01:06:21.460
was super cool and and then somebody took it to a high school but you could still have your you

01:06:21.460 --> 01:06:27.060
know if you were really hyper focused on something if you really were interested in making something

01:06:27.060 --> 01:06:31.140
cool out of coronavirus is because somebody said that something cool could be made out of

01:06:31.140 --> 01:06:35.940
coronaviruses and you're a clever guy maybe that's who Robert over there that's who Ralph

01:06:35.940 --> 01:06:44.660
Berwick is. That's why he's trying to attenuate them. That's why he's trying to find combinations

01:06:44.660 --> 01:06:49.220
of monoclonal antibodies that will neutralize all of them.

01:06:52.580 --> 01:06:57.780
That's why he's trying to change their translation regulatory sequences so that they can't recombine

01:06:58.660 --> 01:07:07.060
with with natural viruses and that they can be permanently attenuated. These are all laudable

01:07:07.060 --> 01:07:14.180
pursuits if viruses are real and and if they are potentially a biotechnology that could be harnessed

01:07:14.180 --> 01:07:22.420
for good. A endogenous signal that could be that could be used and harnessed as a as a tool.

01:07:22.420 --> 01:07:31.380
Why not? Sounds great. I think Ralph Berwick might be innocent. I think we might be led to believe

01:07:31.380 --> 01:07:35.940
that in this little Scooby-Doo this is the guy we're supposed to unmask. This is the bad guy right

01:07:35.940 --> 01:07:42.900
here. Look at him. I don't think so. How do you develop drugs and therapeutic against an unknown

01:07:42.900 --> 01:07:47.940
from this heterogeneous swarm of variant strain that will actually work against the unknown that

01:07:48.020 --> 01:07:53.700
will emerge in the future? How do you target your scarce resources? We think platform technologies

01:07:53.700 --> 01:07:58.420
like synthetic biology, phylogenomics, structure modeling, high-throughput sequencing really will

01:07:58.420 --> 01:08:04.420
provide a platform and our goal is to use SARS coronavirus and its large pool of zoonotic viruses

01:08:04.420 --> 01:08:09.940
as a model to develop rapid response platforms to protect against this broader heterogeneous

01:08:09.940 --> 01:08:16.020
pool of variants. We also would like to develop strategies that would attenuate all family members

01:08:16.020 --> 01:08:20.580
so that you could rapidly develop vaccines for the public overall public health.

01:08:20.580 --> 01:08:25.300
And so that's funny because he's talking about rapidly developing vaccines. I wonder

01:08:25.300 --> 01:08:31.700
anywhere in this talk he's going to project a time frame. Is he going to say five years?

01:08:33.060 --> 01:08:37.140
Because if he says two years that would be very interesting, wouldn't it? If he says

01:08:37.860 --> 01:08:42.900
five years or he says one year anything that he says about development time will be interesting

01:08:42.980 --> 01:08:49.460
in 2007. This is going to be cool. Keep your ears open. Now a lot of beautiful work in Malek's

01:08:49.460 --> 01:08:55.940
lab as well as other labs here in China worked out the basic molecular epidemiology of the SARS

01:08:55.940 --> 01:09:01.060
coronavirus. The virus originated most likely in bat. It probably jumped into civets or humans

01:09:01.700 --> 01:09:07.780
and in that context set up a transmission cycle between civets and ratoon dogs and marketplaces

01:09:07.780 --> 01:09:13.140
and humans who frequented those marketplaces. Over time it's selected for strains that were

01:09:13.140 --> 01:09:17.540
more efficient at infecting human cells and recognizing the human ACE2 receptor for doctor

01:09:17.540 --> 01:09:24.420
human entry leading to the 2002-2003 epidemic which caused about 8,000 cases and 800 deaths.

01:09:26.500 --> 01:09:30.980
Now I want you to keep 8,000 cases and 800 deaths in mind as the

01:09:31.060 --> 01:09:44.740
as a certain quantity of clone. Let's call that one clone unit and so if you release one clone

01:09:44.740 --> 01:09:56.820
unit in a place in China and it can cover 8,000 trackable or suspected cases and 800 deaths

01:09:57.460 --> 01:10:08.260
with a few sequences. So let's just call that a single clone unit as 8,000 cases and 800 people

01:10:08.260 --> 01:10:16.820
dead so that we can think about what we would do if we had two or three clone units, if we had

01:10:16.820 --> 01:10:25.700
10 clone units released in four places. How many cases would that be? How far would it go?

01:10:26.900 --> 01:10:33.540
How much molecular change would occur per unit time in that scenario? I want you to start

01:10:33.540 --> 01:10:40.020
thinking about the possibility that that's what actually was done. The first time single clone

01:10:40.020 --> 01:10:48.340
unit this time maybe five clone units and the difference is that these five clone units were

01:10:48.340 --> 01:10:55.460
all so clones right they're the same so instead of having a SARS outbreak originate from a apartment

01:10:55.460 --> 01:11:04.820
building in China and ultimately end up somewhere in Toronto of all places which was probably part

01:11:04.820 --> 01:11:09.300
of an exercise again because they were elevated as people who knew what they were talking about

01:11:09.380 --> 01:11:18.420
at the beginning of this one and so with more clone units you could just have a bigger brighter

01:11:18.420 --> 01:11:26.820
signal that would be as homogenous as you wanted it to be and so interestingly enough when we were

01:11:26.820 --> 01:11:37.060
having this argument before Kevin McCurnan and I he brought up the idea that there should be some

01:11:37.060 --> 01:11:44.420
purity in the thing which I find curious and I just want to throw that up there for people who

01:11:44.420 --> 01:11:54.420
are listening and watching so this was from earlier this year something you know the things that he

01:11:54.420 --> 01:12:00.260
says that I ignore this is from a slide that I made for that I think it was in the in the last week

01:12:00.260 --> 01:12:06.180
of April the first week of May ask yourself why he's making six hour scooby-doo videos instead

01:12:06.180 --> 01:12:11.060
of downloading data from NCBI and trying to find a signal for his hypothesis in real data

01:12:11.060 --> 01:12:19.060
if the clones offer some advantage to Dr. Evil surely there is evidence of reduced mutation rates

01:12:19.060 --> 01:12:26.340
to prove this now interestingly I didn't want to make a big deal about this I went really fast

01:12:26.340 --> 01:12:30.820
through it because I'm a little scared it's going to disappear I'm a little scared that somebody's

01:12:30.820 --> 01:12:37.460
going to make an excuse for it to disappear but this is the paper from Alina Chan in May of 2020

01:12:37.940 --> 01:12:46.180
Alina Chan with the book Alina Chan that works at the Broad Institute that Eric Lander is the head

01:12:46.180 --> 01:12:54.420
of at MIT Eric Lander the former boss of Kevin McCurnan at the Human Genome Project ladies and

01:12:54.500 --> 01:13:01.860
gentlemen the one that worked at the White House for Biden yeah him now runs the Broad

01:13:01.860 --> 01:13:06.820
Institute or still runs the Broad Institute at MIT where Alina Chan works Alina Chan submitted

01:13:06.820 --> 01:13:14.820
this paper this pre-print to bio archive on May in 2020 and here she has nucleosides nucleotide

01:13:14.820 --> 01:13:22.820
substitutions in the genome of SARS-1 versus nucleotide substances substitutions in this genome

01:13:22.820 --> 01:13:29.860
of SARS-CoV-2 and lo and behold what is this it looks like there is evidence of reduced mutation

01:13:29.860 --> 01:13:34.660
rates at the beginning of the pandemic relative to the beginning of the pandemic of SARS

01:13:36.580 --> 01:13:45.060
hugely reduced mutation rates publicized and observed by none other than Alina Chan who was

01:13:45.060 --> 01:13:49.780
incidentally has never published this paper officially under peer review

01:13:50.500 --> 01:13:57.300
and instead accepted a position at the Broad Institute as a staff scientist and sold a book

01:13:57.300 --> 01:14:06.020
about the lab leak it's a pretty interestingly tiny circle of monkeys considering the fact

01:14:06.020 --> 01:14:17.300
that Kevin McCurnan just recently said that I gave him the George Webb treatment like I mean

01:14:17.300 --> 01:14:22.500
have we are really running out of characters here is it really that many people aren't backstage

01:14:22.500 --> 01:14:31.060
anymore or what like when we when we spike this football ladies and gentlemen it's going to be

01:14:31.060 --> 01:14:37.380
quite a spike okay it's going to be quite a spike let's go back to some more traditional virology

01:14:37.380 --> 01:14:43.140
with the Great Ralph Barrack if you do phylogenetic analysis of the isolates that were sequenced

01:14:45.620 --> 01:14:52.340
you find they fall into two broad categories these this line indicates the strains underneath

01:14:52.340 --> 01:14:58.020
that line were identified during the epidemic and you have early phase isolates primarily of

01:14:58.020 --> 01:15:04.260
January 2003 after a variety of super spreading events you had middle phase isolates shown here

01:15:04.260 --> 01:15:11.460
by this group of isolates and then following infected physicians who came to Hong Kong and

01:15:11.460 --> 01:15:16.100
resulted in a super spreading event that some transmitted the virus to the rest of the world

01:15:16.100 --> 01:15:22.100
these were considered late phase isolates now if anything is suited to a super spreader event

01:15:22.100 --> 01:15:27.780
it is somebody who is infected with an infectious clone because all their cells will have been

01:15:27.780 --> 01:15:34.020
infected with a genomic RNA that was identical which means when they start making this swarm

01:15:34.020 --> 01:15:42.180
of of slightly altered viral genomes and they make this huge collection of sub genomic RNAs

01:15:42.740 --> 01:15:47.860
the collection of proteins that they produce will be much more homogenous than anything

01:15:47.860 --> 01:15:53.860
that would have been generated from a typical cultured virus if you could culture it but you

01:15:53.860 --> 01:15:58.340
can't you see you see what's magical about this you see how beautiful this is

01:16:01.380 --> 01:16:06.340
and so Ralph knows that he's making something that's super cool Ralph knows that he's making

01:16:06.340 --> 01:16:14.180
something that is not natural but he's arguing that not only can we make something super pure

01:16:15.380 --> 01:16:22.580
but if we find a way to alter it so that it's not dangerous so that it's useful to us we can make

01:16:22.580 --> 01:16:31.060
huge quantities of that useful new synthetic virus from Ralph Barrack's perspective he is on

01:16:31.060 --> 01:16:38.420
the verge of tapping into a biotechnology that has almost limitless possibilities

01:16:41.060 --> 01:16:46.900
once he can figure out a way to attenuate them so that they replicate in a way that we want them

01:16:46.900 --> 01:16:53.140
to that they have no toxic parts to them that the most toxic parts of their replication process

01:16:53.140 --> 01:16:59.860
are minimized then potentially we would have the ultimate transfection tool the ultimate gene

01:16:59.860 --> 01:17:06.980
therapy tool the ultimate personalized medicine tool that's what he sees here that's what Ralph

01:17:06.980 --> 01:17:14.420
Barrack is chasing he's chasing a universal vaccine a universal transfection tool a universal gene

01:17:14.420 --> 01:17:22.580
therapy that's what he's teaching chasing here and he wants to use this diverse set of coronaviruses

01:17:22.580 --> 01:17:29.620
in the wild that's present and sustaining in the wild in these populations has a basis for understanding

01:17:29.620 --> 01:17:39.540
them if this biology is it checks out it's a very noble effort in my humble opinion it's not the

01:17:39.540 --> 01:17:48.820
Scooby-Doo bad guy we're looking for it's just not the vast pool of heterogeneous zoonotic strains

01:17:48.820 --> 01:17:53.540
however reside up here most of these were in fact none of these were ever actually successfully

01:17:53.540 --> 01:18:01.460
cultured they actually exist as sequence signatures in silico so one of the immediate goals that we

01:18:01.460 --> 01:18:08.420
became interested in during the outbreak was to develop a platform of viruses that captured the

01:18:08.500 --> 01:18:14.340
heterogeneous that exists within the family of viruses and to do that we identified sort of

01:18:14.340 --> 01:18:19.140
representative strains for example middle phase early phase isolate or bony we had in the lab

01:18:19.140 --> 01:18:26.740
middle phase isolate with cohk w1 kzo2 with early phase isolate animal isolates like it's from sivets

01:18:26.740 --> 01:18:34.100
fc 16 and hc fc 6103 a raccoon dog isolate which is shown down here the sporadic 2004 human case

01:18:35.060 --> 01:18:41.380
and a couple of other viruses sequences were identified as good candidates that would capture

01:18:41.380 --> 01:18:48.180
all the diversity that had been identified within the sequence pool this shows the sequence that's

01:18:48.180 --> 01:18:54.100
the sequence variation within the spike like a protein if it's a blue box amino acid that's an

01:18:54.100 --> 01:18:59.780
animal associated residue early phase changes are shown here in yellow middle phase changes

01:18:59.780 --> 01:19:05.860
shown in orange and late phase changes shown in red in addition in 2004 there are a variety

01:19:05.860 --> 01:19:10.580
of other animal strains of sporadic human cases were identified that had additional variations

01:19:10.580 --> 01:19:16.820
shown here in white and it's interesting as much i i hope that everybody was paying attention right

01:19:16.820 --> 01:19:25.620
what i put up here before you were paying attention right um this paper is actually

01:19:26.580 --> 01:19:32.740
this paper this paper right here

01:19:36.740 --> 01:19:42.660
broad spectrum coronavirus antiviral drug discovery by allicin totura and cena bivari

01:19:42.660 --> 01:19:48.420
written in 2019 allicin totura was a postdoc of ralph veric

01:19:48.420 --> 01:19:55.620
so she's at us amrit

01:19:59.140 --> 01:20:01.620
you don't think she knows how to make infectious clones

01:20:03.620 --> 01:20:07.300
you don't think everybody that went through ralph veric's lab knows how to make infectious

01:20:07.300 --> 01:20:13.860
clones are you kidding me where did these people go off to they're not working at starbucks or

01:20:13.860 --> 01:20:16.980
something like that you think

01:20:20.500 --> 01:20:25.460
this paper is not accidental that it talks about a virus getting out of china and needing to use

01:20:25.460 --> 01:20:32.900
remdesivir as an antiviral for it it's not for nothing that we're defending the proof reading

01:20:32.900 --> 01:20:41.460
ability of exo x one gene or or our exo and gene uh the the ribonuclea we're not we're not

01:20:41.460 --> 01:20:49.700
defending these these things for nothing they're all related all these people all their nonsense

01:20:49.700 --> 01:20:58.740
it's all related don't you remember the the the domain program run by ditra

01:21:00.340 --> 01:21:06.820
run by robert malone who supposedly made an x-ray crystallography computer model

01:21:06.900 --> 01:21:15.060
of the three cl protease of the virus and then interface that computer model with all known

01:21:15.060 --> 01:21:20.500
pharmaceuticals with a volunteer team of researchers in a matter of weeks to identify

01:21:20.500 --> 01:21:29.300
femotidine and remdesivir as two usefully useful antivirals

01:21:37.460 --> 01:21:40.020
and cena bivari knew about it a year before that

01:21:41.700 --> 01:21:45.620
and ralph veric and mark dennison knew about it a few years before that

01:21:46.580 --> 01:21:51.220
for zika or for for Ebola for whatever else they were trying it on

01:21:56.500 --> 01:21:58.740
because they're all RNA viruses right

01:22:00.980 --> 01:22:06.660
don't you see the pattern here there is a mythology that has been laid down there is a

01:22:06.660 --> 01:22:08.500
mythology that is being defended

01:22:08.900 --> 01:22:20.580
this paper got in bobbie's book this paper that is in bobbie's book and actually actually

01:22:20.580 --> 01:22:26.500
right uh uh robert malone sold bobbie's book today on his sub stack which is pretty badass i like

01:22:26.500 --> 01:22:33.460
that a lot maybe that was part of the deal if you get that you stop that kooie from talking about

01:22:33.540 --> 01:22:40.580
me on his stream i'll i'll sell bobbie's book on my sub stack maybe that's what it is

01:22:41.780 --> 01:22:46.260
it's a pretty simple deal right you know robert malone's got a million subscribers if he tweets

01:22:46.260 --> 01:22:49.860
out a book probably a hundred thousand people buy it or ten thousand people buy it

01:22:52.660 --> 01:22:57.540
he better sub stack the book he wrote a wrote a review on the back cover

01:22:58.420 --> 01:23:06.900
i'd say buy a copy i'd say buy a copy and we'll meet and i'll autograph it for you

01:23:07.860 --> 01:23:12.820
um there's some bad stuff in there there's some good stuff in there it's probably the best

01:23:12.820 --> 01:23:14.820
document of all the nonsense people said

01:23:18.580 --> 01:23:22.980
the person who showed me this paper the person who showed me this paper his Mark

01:23:23.540 --> 01:23:29.780
Housatonic Mark Kulacz it wasn't shown to me by by Kevin McCairn it wasn't shown to be by

01:23:29.780 --> 01:23:36.580
Paul Cottrell it wasn't shown to me by George Webb wasn't shown to me by Kevin McKernan or Stephanie

01:23:36.580 --> 01:23:44.580
Sinev or Steve Kirsch or Robert Malone the only person that's ever mentioned this paper to me

01:23:44.580 --> 01:23:48.740
ever shown it to me ever found it and said holy crap dude did you read this

01:23:49.300 --> 01:23:51.780
this is Mark Kulacz

01:23:55.780 --> 01:24:00.900
i just can't stress to you how all of these things line up in a way that that can't be

01:24:00.900 --> 01:24:06.340
discounted anymore all of these people all of their behavior line up in a way that cannot

01:24:06.340 --> 01:24:11.300
be discounted anymore even if we find out that there's a huge coronavirus background that

01:24:11.300 --> 01:24:18.180
coronaviruses can transmit for months even if we find out that clones can transmit for months

01:24:19.620 --> 01:24:26.580
the ultimate conclusion is is that we have been lied to by a group of medlers that sustained

01:24:26.580 --> 01:24:34.980
a worst-case scenario sustained a confusion and uncertainty and a doubt that allowed the world

01:24:34.980 --> 01:24:41.620
to be transfected and only after the world was transfected that these people do it about face

01:24:41.620 --> 01:24:47.940
and pretend like they always knew and the list of these people is long but it doesn't include

01:24:47.940 --> 01:24:55.460
Ralph Baric the list of people who lied to us about the worst-case scenario at the beginning

01:24:55.460 --> 01:25:02.020
of the pandemic only to pivot after our kids had lost two years of school after our kids have

01:25:02.020 --> 01:25:09.220
been traumatized with masks after we had been locked down for a year and a half that's not Ralph Barrick

01:25:09.220 --> 01:25:21.380
case you can't tell i'm trying to get Ralph Baric to come and interview on Gigaohm

01:25:21.380 --> 01:25:27.220
Biological wouldn't that be badass much of the variation actually falls within residues or regions

01:25:27.220 --> 01:25:33.620
that neutralizing epitopes were identified and in fact although the number of residues that are

01:25:33.620 --> 01:25:38.420
changed aren't that great you can actually reduce neutralization titers by about 20 fold

01:25:39.460 --> 01:25:47.300
with some of these variants now importantly Farzan's group showed that the two key changes

01:25:47.300 --> 01:25:52.500
during the epidemic where this life seemed to asparaging change at 4.79 and Assyrian 3

01:25:52.500 --> 01:26:00.340
change at 4.87 that drove animal adaptation to human strain and so keep that in mind so

01:26:01.060 --> 01:26:10.340
without access to this pool of strains we decided to synthesize basically a portfolio of

01:26:11.060 --> 01:26:16.980
spike like a protein genes of about 4k each and then use a molecular clone for the

01:26:16.980 --> 01:26:22.100
urbani epidemic strain that we had built in the lab basically replacing the urbani spike

01:26:22.100 --> 01:26:26.020
one at a time with these various spike like a protein from different phases of the epidemic

01:26:26.820 --> 01:26:32.580
now all of these viruses were viable except for the sc-16 variant this actually can't recognize

01:26:32.580 --> 01:26:38.100
the humanase receptor so it didn't grow until we made mouth cells that consistently expressed

01:26:38.100 --> 01:26:42.980
the civetase to receptor and once we did that this virus to grow well before we had those cells

01:26:43.620 --> 01:26:50.260
since Farzan had identified this 4.79 changes the key residues so be careful there now they

01:26:50.260 --> 01:26:55.620
just admitted that they use mouth cells which have been genetically altered to express the

01:26:55.620 --> 01:27:02.900
civetase receptor now i can't stress enough we had this paper come out that everybody's been

01:27:02.900 --> 01:27:07.460
yelling about it i'm probably gonna have to do a show about it in the next couple days again

01:27:08.260 --> 01:27:14.980
about this paper that um you know i should do that video tomorrow that's what i'm gonna do

01:27:14.980 --> 01:27:24.980
tomorrow i'm gonna do i found this video with mccernan and um christine uh parks christina parks

01:27:25.860 --> 01:27:35.140
and um John Beaudoin and i've i've been meeting to do John Beaudoin um on HighWire and so I should

01:27:35.140 --> 01:27:41.860
do John Beaudoin on HighWire this week and then I'll do this this multiple person stream that

01:27:41.940 --> 01:27:46.740
happened a few days a few a week ago or so with McKernan and parks and Beaudoin that'll be good

01:27:46.740 --> 01:27:52.900
that's two good good study halls for this week um so what he's talking about here now is making a

01:27:52.900 --> 01:27:57.940
number of different clones where he's swapping the spike out um this is of course supposed to get

01:27:57.940 --> 01:28:03.300
us really excited about making chimeric viruses but here again he's gonna make clones of all these

01:28:03.300 --> 01:28:09.700
things so if there's a danger the danger is is that he's making clones if he wasn't making clones he

01:28:09.700 --> 01:28:16.740
couldn't even study these things the regular virus of these things is not attainable won't grow

01:28:16.740 --> 01:28:21.940
and even if you could grow it the titers would be so low it would be almost unsequitable

01:28:23.300 --> 01:28:30.980
that's the problem with these guys right and so don't let the the nonsense fool you

01:28:30.980 --> 01:28:38.420
infectious clones are the bedrock methodology of RNA virology change especially build a recombinant

01:28:38.420 --> 01:28:44.180
virus with that change and that virus could be cultured uh interestingly enough two of these

01:28:44.180 --> 01:28:50.340
the gz-o-2 and the uh hcse-61 of three viruses actually caused lethal infection in aged animals

01:28:50.340 --> 01:28:56.020
they produced an ARDS like disease saw SARS caused ARDS in humans or predominantly in age

01:28:56.020 --> 01:29:01.300
populations uh that's acute respiratory distress syndrome it's a clinically devastating end stage

01:29:01.300 --> 01:29:06.420
lung disease with about a 50 mortality rate so these are actually some of the first real good

01:29:06.420 --> 01:29:13.300
animal models for SARS infection and so remember what I said the other day again you have to

01:29:14.100 --> 01:29:19.380
take everything with a huge grain of salt if it's an animal model especially a genetically modified

01:29:19.380 --> 01:29:27.700
one then the expression of the protein that the virus interacts with could be so over or so

01:29:27.780 --> 01:29:39.140
inappropriate or any other number of of wrong that when exposed to these exosome collections

01:29:39.140 --> 01:29:44.740
these animals produce a really robust immune response or a really robust infection because

01:29:45.300 --> 01:29:49.780
these signals go everywhere so if you believe the biology that they need a receptor and that

01:29:49.780 --> 01:29:54.420
the receptor has to be somewhere and if the receptor isn't there then the virus can't hurt you

01:29:54.980 --> 01:30:01.700
if you believe this then these animal models are also very scary because you could imagine

01:30:01.700 --> 01:30:07.300
a scenario whereby genetically altering the animal you create an animal that is hyper

01:30:07.940 --> 01:30:14.740
susceptible to this manipulation hyper susceptible to the exposure to these signals

01:30:15.700 --> 01:30:22.740
so if you believe viruses work the way that these people say they do and that they need receptors

01:30:22.820 --> 01:30:27.940
and then you alter the expression of those receptors or even worse you homogenize them

01:30:27.940 --> 01:30:34.180
across the whole animal then their susceptibility to exposure to these things could be very different

01:30:34.180 --> 01:30:40.180
and that that could be great if you're writing grant applications that need an animal model that's

01:30:40.180 --> 01:30:46.900
very repeatable and very robust and so don't underestimate the stuff that we talked about with

01:30:47.460 --> 01:30:53.620
with Wolfgang Wodock keeps coming up here where you have academic biologists that are essentially

01:30:53.620 --> 01:31:01.060
academy-gicians they've been they become so good at doing experiments that are fundable

01:31:02.020 --> 01:31:08.980
asking questions that are fundable that they have stopped asking useful questions that they have

01:31:08.980 --> 01:31:14.900
stopped asking questions that actually move the ball forward but they're now asking sort of circular

01:31:14.980 --> 01:31:22.580
questions they don't want to make any progress they want to make the illusion of motion right

01:31:24.420 --> 01:31:29.140
and so in this scenario i don't think that's what what Ralph barracks doing at all i think he's

01:31:29.140 --> 01:31:35.460
actually making forward progress there are these RNA signals which have been very difficult to study

01:31:35.460 --> 01:31:41.940
in the wild because they're almost unculturable there's no infectious material available it's so

01:31:41.940 --> 01:31:47.300
rare and if you take this infectious material and try to culture it you often get very low to

01:31:47.300 --> 01:31:58.260
no tighter so infectious clone overcomes that Ralph barrack is a genius now um these viruses

01:31:58.260 --> 01:32:03.780
replicate of the human epidemic strains replicate efficiently in human airway epithelial cultures

01:32:03.780 --> 01:32:08.500
these are cultures that are derived from cells lining the trachea of transplant patients you can

01:32:08.580 --> 01:32:13.780
actually culture them on liquid air interfaces and they take on the architecture of the human airway

01:32:13.780 --> 01:32:20.420
stars coronavirus all the epidemic strains actually like to infect ciliated cells and these epidemic

01:32:20.420 --> 01:32:26.900
strains replicate very efficiently the animal strains however do not and this is just some

01:32:26.900 --> 01:32:33.540
fluorescent microscope images showing the cilia of the ciliated cells with an expression of the

01:32:33.540 --> 01:32:39.220
sars nucleic acid protein from an epidemic strain on the ciliated cells and the zoonotic

01:32:39.220 --> 01:32:47.140
strains se 16 in this and hdse 6103 and replicate however if you passage these viruses on human

01:32:47.140 --> 01:32:52.660
airway cells and culture you can actually rapidly select out variants that can replicate efficiently

01:32:52.660 --> 01:32:59.540
in human airways those viruses actually do not contain the mutations that would be had been

01:32:59.620 --> 01:33:05.780
predicted by mike farson as being which were clearly responsible for the 2002-2003 epidemic

01:33:06.340 --> 01:33:14.100
rather we saw changes um at positions 442 and 472 and 479 and so this is an interesting

01:33:14.100 --> 01:33:23.300
presentation of the data where it seems like this single trial of putting a clone on a epithelial

01:33:23.300 --> 01:33:30.100
cell culture a differentiated epithelial cell culture and then seeing sequence changes in the

01:33:30.740 --> 01:33:37.860
consensus sequence he seems to talk about it like okay so we put the car on the road and it drove

01:33:37.860 --> 01:33:42.180
down the road and then that's how it works and so if we did it again that's what would happen

01:33:43.380 --> 01:33:49.300
and i don't think that that's really how virology works i think if he did this again he would get

01:33:49.300 --> 01:33:57.300
different results here it might be a similar kind of selection process by which non-infectious

01:33:57.300 --> 01:34:04.580
particles versus infectious particles are selected and and can be seen in in in subsequent passages

01:34:04.580 --> 01:34:10.660
or detected in subsequent passages but that selection process is purely based on function

01:34:10.660 --> 01:34:18.020
then it's not going to be based on fitness nothing is is fitness unless it's number of copies right

01:34:18.020 --> 01:34:23.300
so then we're talking about a whole different thing and we're really only doing a selection

01:34:23.300 --> 01:34:28.340
process we're taking the whole supernatant here there's there's no like you know

01:34:29.540 --> 01:34:34.740
race to see who can make more copies of themselves we're not selecting based on fidelity or anything

01:34:34.740 --> 01:34:38.980
like that in fact we have the argument that there has to be mutation rate otherwise there would be

01:34:39.060 --> 01:34:46.660
a collapse so it's interesting um that's it's that's it mediated the cross species transmission

01:34:46.660 --> 01:34:52.500
event so in reality we've done this a couple of times there's actually several pathways by which

01:34:52.500 --> 01:34:57.940
the zoonautics are it could actually adapt and recognize the humanase receptor what's interesting

01:34:57.940 --> 01:35:04.340
is that the epidemic strains actually efficiently use both the humanase receptor and the civetase

01:35:04.420 --> 01:35:10.260
receptor when we in vitro select for human adapted strains on human airway culture

01:35:11.140 --> 01:35:16.820
they actually lose the ability to recognize the civic receptor so what this data suggests

01:35:16.820 --> 01:35:22.900
actually is that ours had existed in a transmission cycle between humans and civets actually probably

01:35:22.900 --> 01:35:28.500
for several several years prior to the 2002-2003 epidemic allowing the virus to

01:35:28.580 --> 01:35:34.900
regain the capacity to recognize both receptors okay so those were the easy ones to do

01:35:35.860 --> 01:35:41.700
oh with an application um so now that we had a large panel of our of uh variant viruses we could

01:35:41.700 --> 01:35:46.500
use those for therapeutic testing over vaccines in this case i'm going to show you an example of

01:35:46.500 --> 01:35:52.420
about 30 human monoclonal antibodies that were derived by Antonio Lanzavecchia if you take those

01:35:52.420 --> 01:35:56.820
30 monoclonals and test their ability to neutralize all the strains that we've made in the laboratory

01:35:56.820 --> 01:36:03.220
you find some that only neutralize urbani some that neutralize all human strains some that neutralize

01:36:03.220 --> 01:36:08.820
some civets but not all the animal strain but you do end up with four antibodies that neutralize

01:36:08.820 --> 01:36:14.340
all strains we have in our portfolio including the ones that were in vitro adapted on human

01:36:14.340 --> 01:36:22.260
airway culture importantly if you select for escape using these antibodies these antibodies

01:36:22.260 --> 01:36:26.420
select for changes in different locations of the receptor binding domain

01:36:26.420 --> 01:36:32.020
and in fact three of them actually don't overlap yes 109 the 230 and the 227

01:36:32.020 --> 01:36:37.380
don't overlap and so these represent good therapeutic cocktails that would capture most of the

01:36:37.380 --> 01:36:43.460
diversity that exists in SARS therapeutic cocktail what an interesting story we can tell there so

01:36:44.260 --> 01:36:51.380
there's a guy by the name of plumber who used to work on AIDS in Canada who died at the beginning

01:36:51.460 --> 01:37:03.140
of the pandemic he had a postdoc I think she was Chinese and she had a three antibody cocktail

01:37:03.140 --> 01:37:13.620
called Z map that was trialed against remdesivir and was actually being trialed against remdesivir

01:37:13.620 --> 01:37:20.100
probably at us amrit right before the pandemic and then it was shut down

01:37:21.700 --> 01:37:30.020
so the trial comparing remdesivir with Z map was stopped this is all Kevin sorry this is all

01:37:30.020 --> 01:37:39.140
marcoolax work here that I'm reciting from memory so we have an AIDS expert dying I think he fell

01:37:39.140 --> 01:37:46.980
on the sidewalk or something crazy like that and we have this spy Chinese lady who supposedly

01:37:48.100 --> 01:37:53.700
took some samples back to China and so she's a spy she she got put away but actually she was

01:37:54.260 --> 01:38:03.540
like on the cover of magazines and stuff is being the star of of of biology one year for Z map it's

01:38:03.540 --> 01:38:11.460
really cool because that was a three antibody cocktail this is also another proposed three

01:38:11.460 --> 01:38:21.220
antibody cocktail as a as a pan by an antiviral that's pretty that's pretty cool I I'm starting

01:38:21.220 --> 01:38:28.020
to see Ralph Barrett as a good guy it's weird and probably work in potential patients that would

01:38:28.100 --> 01:38:33.140
be infected during future epidemics and these also actually protect young and aged animals from

01:38:33.140 --> 01:38:40.260
lethal infection that was a paper published by Barry Rock in a couple papers now the civet

01:38:40.260 --> 01:38:46.420
and the raccoon dog strains were the easy ones the true reservoir for SARS is within bat population

01:38:47.460 --> 01:38:52.980
so if you look at a phylogenetic tree the human strains are shown here in red the civet strains

01:38:52.980 --> 01:38:57.540
raccoon dog strains are shown in purple but the real variation is within the bat strain

01:38:58.580 --> 01:39:02.340
um the reservoir it's thought that these were probably the reservoir for the

01:39:03.140 --> 01:39:09.220
emergence of the virus now these are about 80 to 90 percent identical to SARS they can't be

01:39:09.220 --> 01:39:14.100
cultured and they exist to sequence signature signatures in silico there's also extensive

01:39:14.100 --> 01:39:20.020
variation see they can't be cultured they exist as sequence signatures in silico well that just

01:39:20.020 --> 01:39:26.900
means they're stored on a computer they're keyboard viruses and so clones are really really important

01:39:26.900 --> 01:39:33.940
because you can take a virus that you've only detected in the wild as a sequence and create

01:39:33.940 --> 01:39:41.060
large quantities of that RNA and electropyrated into cells and get those cells to run it through

01:39:41.060 --> 01:39:49.460
their ribosomes and package it the only thing you can't do sometimes is passage it

01:39:49.940 --> 01:39:57.940
but the cells can be made to take the RNA up they can be it can be electropyrated in

01:40:00.020 --> 01:40:05.540
you can use lip effectamine to get it in and then those cells will read the RNA and those

01:40:05.540 --> 01:40:09.780
proteins will produce and the sub genomic RNAs will be produced and everything will be packaged

01:40:09.860 --> 01:40:20.740
up and then you'll get this this effect this is cool because this is really underscoring

01:40:22.580 --> 01:40:29.700
that downplaying the clones is just it's absurd it's absurd it is a bedrock methodology of RNA

01:40:29.700 --> 01:40:34.980
virology through the replicase and elsewhere in the genome so when we decided we were going to

01:40:34.980 --> 01:40:41.620
synthetically resurrect the entire virus now before we started it's important to note this is a

01:40:44.340 --> 01:40:51.220
the cryoem reconstruction of the SARS glycoprotein spike notice that there are three receptor binding

01:40:51.220 --> 01:40:57.300
domains shown here in with bubbles that actually engage the ACE2 receptor if you look at the sequence

01:40:57.300 --> 01:41:03.860
the receptor binding domain shown here in yellow there's a large amount of sequence variation

01:41:03.860 --> 01:41:09.060
especially within the contact interface residues that engage the human ACE receptor

01:41:09.060 --> 01:41:13.620
and in fact there's only four of 13 contact interface residues that are retained in these

01:41:13.620 --> 01:41:18.580
fat strains but we don't actually think it's going to use the human ACE receptor to get into cells

01:41:19.300 --> 01:41:24.580
in fact we think we're going to have a tough time culturing the virus now it's important to note

01:41:24.580 --> 01:41:31.700
that the SARS RBD that is was identified has been proposed by a couple of groups that it may have

01:41:31.700 --> 01:41:37.220
been introduced by recombination processes from unknown strains that haven't yet been identified

01:41:37.220 --> 01:41:42.820
and that that led to the initial cross-species transmission events now to get back to the

01:41:42.820 --> 01:41:48.900
issue of in silico sequences they actually represent hypothetical viruses most synthetic viruses that

01:41:48.900 --> 01:41:55.540
have been resurrected to this point actually we knew that the sequence was infectious in this case

01:41:55.540 --> 01:42:01.460
we actually don't know which of the sequences in GenBank were infectious if any so to do that

01:42:01.460 --> 01:42:05.860
with an error rate in GenBank ranging from about one to five hundred to one to ten thousand depending

01:42:05.860 --> 01:42:12.100
on the sequence we had to do extensive bioinformatic analysis to identify what we thought was the likely

01:42:12.100 --> 01:42:18.020
consensus sequence well that sounds like that sequencing is a lot more hairy than i thought it was

01:42:18.740 --> 01:42:22.980
I just thought sequences were sequences you get one it just kind of spits it out

01:42:22.980 --> 01:42:26.900
beep beep beep beep beep beep beep beep beep beep beep beep beep beep beep beep beep beep beep

01:42:26.900 --> 01:42:32.660
then you just kind of read it off right now we're getting back to the Vincent Rancin yellow

01:42:32.660 --> 01:42:41.940
explanation of consensus sequence the the more traditional Vincent Rancin yellow will say that

01:42:41.940 --> 01:42:50.820
the consensus sequence might not even actually exist but Kevin McCurnan says that there are 15

01:42:50.820 --> 01:42:57.380
million SARS-CoV-2 genomes on on gents said they're all real they're all independent they all come

01:42:57.380 --> 01:43:03.140
from different people so they have to be real what's Ralph Barrick talking about here then

01:43:03.940 --> 01:43:16.500
I see a battleship size incongruency between pre-COVID virology and post-COVID virology

01:43:19.860 --> 01:43:25.300
I really do I really think we're on to something here I think with a couple weeks of work

01:43:25.940 --> 01:43:34.260
this is going to be over and we're going to be able to you know incorporate all of these

01:43:34.260 --> 01:43:40.500
various factions into a common explanation for why a lot of people were mistaken

01:43:41.460 --> 01:43:47.700
why a lot of people were fooled why you could be taken all the way by no virus why you could

01:43:47.700 --> 01:43:52.900
be taken all the way by lab leak why you could be taken all the way by my market zoneosis and

01:43:52.980 --> 01:43:58.660
why you could be taken all the way by by any of these other in-betweens including

01:44:00.100 --> 01:44:01.700
including iatrogenic murder

01:44:04.420 --> 01:44:11.460
but in the end there's only one explanation that fits all of them that lets the most people on

01:44:11.460 --> 01:44:22.660
earth be right and I think that's why everybody's so angry because the explanation that we are being

01:44:22.660 --> 01:44:28.900
fooled into believing it's Ralph Barrick and Tony Fauci and eco-health alliance

01:44:30.420 --> 01:44:39.940
instead of understanding that this technology was actually available available it was available

01:44:40.900 --> 01:44:41.860
to us Amrit

01:44:44.420 --> 01:44:50.020
probably for a couple decades right ever since they first did it with the polio virus

01:44:53.540 --> 01:44:57.860
we have to wake up and apologize to our kids sooner or later we might as well do it now

01:44:58.660 --> 01:45:03.380
none of the strains that we actually saw we thought was completely correct some of them had

01:45:03.380 --> 01:45:08.980
deletions in the sequence of the five prime ends that we had to make some educated guess

01:45:10.180 --> 01:45:14.900
the basic approach that we build coronavirus is using our molecular clone is shown here

01:45:14.900 --> 01:45:21.860
with the SARS clone shown in blue is the clone is broken into six five kv about five kv thesis

01:45:22.660 --> 01:45:29.620
each piece is flanked by bagel one restriction endonucleate sites these are class two S restriction

01:45:29.620 --> 01:45:35.060
enzymes that recognize a palindromic sequence and so this is asymmetric and this is almost

01:45:35.060 --> 01:45:40.020
certainly one of the enzymes that was mentioned in the foyer request that was recently released

01:45:41.300 --> 01:45:50.580
and so the release of this enzyme or related enzymes as a list in an email or a supplementary

01:45:50.580 --> 01:45:56.580
document or something like that is now being misconstrued as evidence that the diffuse proposal

01:45:56.580 --> 01:46:01.060
is real and that they were making clones and that they were spraying them into bad caves and they

01:46:01.060 --> 01:46:08.980
put fear and cleavage sites at the joint between S1 and S2 yada yada yada so most likely that's

01:46:08.980 --> 01:46:15.780
why we're being fed this video is so that we get to this point right here and we see how clones

01:46:15.860 --> 01:46:24.420
are assembled and now i'm sure he's about to tell us that this right here represents a S2 S1 S2

01:46:26.340 --> 01:46:32.420
clone junction there so that they can substitute the S1 from other coronavirus is right in there

01:46:32.420 --> 01:46:40.260
let's listen this allows since these ends are asymmetric they actually will not concatomerize

01:46:40.260 --> 01:46:44.580
like classic sticky ends left by restriction enzymes but rather they become directional

01:46:45.220 --> 01:46:50.660
so if you end clone A with a bagel site that leaves one three nucleotide overhags and the

01:46:50.660 --> 01:46:55.060
five prime end of the B fragment with the complementary three nucleotide overhags they

01:46:55.060 --> 01:46:59.780
totally get it directly you change different bagel sites at each piece it's midnight

01:47:00.740 --> 01:47:07.940
assemble up into 30k B chromosomes like tinker toys like tinker toys like tinker toys

01:47:07.940 --> 01:47:13.300
batching home that we made that's actually a good way to date somebody like do you know

01:47:13.300 --> 01:47:19.140
what tinker toys man i had tinker toys you know i had lots of them like a lot of them i had a

01:47:19.140 --> 01:47:25.300
lot of tinker toys i think i had like three of the cylindrical containers i could go down

01:47:25.300 --> 01:47:30.740
with tinker toys hey by the way it's uh past midnight so technically speaking i just turned

01:47:30.740 --> 01:47:39.860
52 years old yeah 52 52 years old how's that for old using blue heron and biobasic basically

01:47:39.860 --> 01:47:45.060
is interchangeable with the urbanic clone the only real difference was that we broke the F

01:47:45.060 --> 01:47:50.180
fragment into two pieces so that we could play with the receptor binding domain easily if this

01:47:50.180 --> 01:47:56.020
thing didn't turn out to be infectious and in fact we made this clone we built the fool on cDNA we

01:47:56.020 --> 01:48:00.660
drove transcripts electroprated that in the cells and we can see evidence of replication by

01:48:00.660 --> 01:48:05.540
the synthesis of sub-genomic messenger RNA but we couldn't culture the virus and we couldn't pass

01:48:05.540 --> 01:48:10.420
it from cell to cell ah we couldn't culture the virus we couldn't passage it from cell to cell so

01:48:10.420 --> 01:48:17.460
they could see that the RNA was being sorted into sub-genomic RNAs but they couldn't passage it from

01:48:17.460 --> 01:48:23.620
cell to cell does that mean it was not getting packaged or does that mean that it wasn't binding

01:48:23.620 --> 01:48:28.980
in the next passage i think it wasn't binding in the next passage is the right explanation

01:48:28.980 --> 01:48:35.940
which means that you could make virus from these clones you just have to have a receptive cell type

01:48:35.940 --> 01:48:40.900
on the other end that's going to do it so they transfected listen carefully they transfected

01:48:41.540 --> 01:48:49.460
this clone into that cell culture and it packaged virus for them you know it packaged virus because

01:48:49.460 --> 01:48:54.420
it made the sub-genomic RNAs those were translated into the many many many copies of the proteins

01:48:54.420 --> 01:48:59.860
and all this stuff happened it's just that when they took the supernatant off of that cell culture

01:48:59.860 --> 01:49:06.180
and tried to put it on the next cell culture it didn't propagate and that has to do with the spike

01:49:06.180 --> 01:49:11.860
protein receptor binding domain compatibility with the cell culture that you're using according to

01:49:11.860 --> 01:49:18.020
their methodology explanation so let's listen to that again because i want you to hear what he did

01:49:18.100 --> 01:49:22.900
he electroporated it into a cell culture but couldn't get it to jump from cell culture to cell

01:49:22.900 --> 01:49:30.820
culture which is what culturing is what passaging is clearly there was RNA the cell and we could

01:49:30.820 --> 01:49:36.100
clone we built the full-on cDNA we drove transcripts electroprated that in the cell and we can see

01:49:36.100 --> 01:49:40.740
evidence of replication by the synthesis of sub-genomic messenger RNA but we couldn't

01:49:40.740 --> 01:49:45.380
culture the virus and we couldn't pass it from cell to cell sub-genomic electroprated that and in fact

01:49:45.380 --> 01:49:50.660
we made this clone we built the full-on cDNA we drove transcripts electroprated that in the cell

01:49:50.660 --> 01:49:55.380
and we can see evidence of replication by the synthesis of sub-genomic messenger RNA

01:49:55.380 --> 01:49:59.780
but we couldn't culture the virus and we couldn't pass it from cell to cell so clearly there was

01:49:59.780 --> 01:50:08.740
probably a defect in entry to solve that problem we use literature data that has suggested that

01:50:08.740 --> 01:50:14.020
RBD domains of coronaviruses may be interchangeable between species so we took the human

01:50:14.900 --> 01:50:20.580
the urbani epidemic receptor-bonding domain that's 210 amino acids and went

01:50:21.620 --> 01:50:28.260
and dropped that into the bat genome backbone producing a chimera with the receptor-bonding domain

01:50:28.260 --> 01:50:33.940
driven from the epidemic strains now when we built that clone drove transcripts and electroprated

01:50:33.940 --> 01:50:39.140
that in the cells we got a virus that could replicate quite efficiently this is just some

01:50:39.140 --> 01:50:46.020
growth curve data showing I think the black box is the urbani wild type and the white circles are

01:50:46.020 --> 01:50:52.980
open open symbols are the bat viruses you can see they replicate exactly as good as the urbani

01:50:52.980 --> 01:50:58.900
epidemic strain at low and high multiplicities of infection they recognize the human ACE2 receptor

01:50:58.900 --> 01:51:04.420
and dbt cells at low and high multiplicity infections and grow just like the epidemic strains

01:51:04.420 --> 01:51:10.020
and they also retain the ability to use the CIVID ACE2 receptor just like the epidemic strains

01:51:10.020 --> 01:51:16.500
low and high multiplicity they also are capable of infecting human airway epithelial cultures

01:51:16.500 --> 01:51:21.380
and targeting ciliated cells just like the epidemic strain and they grow to similar titer

01:51:23.700 --> 01:51:28.740
now if you make aniseera just against the bat spike like a protein

01:51:29.620 --> 01:51:36.500
and use that aniseera to target the SARS wild type virus it will not neutralize that virus

01:51:37.220 --> 01:51:43.140
so clearly aniseera and vaccines derived against epidemic strains are not going to protect against

01:51:43.140 --> 01:51:49.540
the bat strains to use that same sera against the RBD chimeric virus you can neutralize it

01:51:49.540 --> 01:51:55.220
indicating that there are neutralizing episodes that resist that exist outside the RBDs and if

01:51:55.220 --> 01:52:00.340
you take the human monoclonal to target the epidemic strain RBDs you can neutralize these

01:52:00.340 --> 01:52:05.700
virus with quite efficient so one thing that I think we all need to learn is exactly what neutralized

01:52:05.700 --> 01:52:13.460
means in this context because I kind of was under the naive operation or operating under the naive

01:52:13.460 --> 01:52:20.580
assumption that that neutralized means to bind to the receptor binding domains so that the receptor

01:52:20.580 --> 01:52:24.580
binding domain is not able to interact with the receptor and therefore you don't get

01:52:25.540 --> 01:52:30.420
infection so I thought that's what neutralizing was but I think neutralizing has more to do with

01:52:30.420 --> 01:52:38.900
like precipitation and if you get enough antibodies stuck onto you then you come out of out of solution

01:52:38.900 --> 01:52:46.020
and I'm wondering if it's something like this too like uh aggregation or something like that

01:52:46.020 --> 01:52:51.460
that causes neutralization and precipitation so I need to learn that a little bit better because

01:52:51.460 --> 01:52:57.860
I do know that that people like Mark Bailey and and uh Steven Lanka have been working on trying to

01:52:57.860 --> 01:53:02.900
understand what it is that virologists are meaning with that and I do think that that is something

01:53:02.900 --> 01:53:11.220
that we should be masters of here as well so I apologize for that so in summary um certainly

01:53:11.220 --> 01:53:15.780
synthetic genomics and reverse genetics this can be a platform to recover uncultivatable

01:53:15.780 --> 01:53:20.580
zoonotic precursors can be used to synthesize this is actually the largest synthetic virus

01:53:20.580 --> 01:53:27.540
to date it's going to be a short record but it is a record um and you can use it now to identify

01:53:27.540 --> 01:53:33.860
a broad spectrum antiviral from the vaccine clearly the RBDs are interchangeable uh this is with an

01:53:33.860 --> 01:53:40.980
89 percent amino acid identity within the spike it's the minimal domain required to host shift

01:53:40.980 --> 01:53:46.420
coronaviruses what is the phylogenetic limits to RBD interchange we actually don't know

01:53:47.620 --> 01:53:52.580
clearly the human monoclonals and vaccines targeting the SARS RBD would provide protective

01:53:52.580 --> 01:53:56.740
immunity against natural isolates that emerge that's not my sound the overall process is in

01:53:56.740 --> 01:54:04.500
future or unfortunately by deliberate design so now I want to move to the next part of the

01:54:04.500 --> 01:54:09.700
talk which is synthetic the optimization scheme to attenuate SARS coronavirus pathogenesis

01:54:10.580 --> 01:54:16.420
this is the maturation pathway okay so after as he explains the maturation pathway I want you to

01:54:16.980 --> 01:54:22.340
imagine that we're trying to solve the puzzle of the infectious cycle as well and then I also

01:54:22.340 --> 01:54:30.820
want you to listen carefully to the amount of effort time thought and ideas that are going into

01:54:31.620 --> 01:54:42.340
the attenuation of coronaviruses and a general strategy to attenuate all coronaviruses specifically

01:54:42.340 --> 01:54:50.580
in the context of recombination and so he's keenly aware that there's a problem with the

01:54:50.580 --> 01:54:56.100
oral poliovirus vaccine that when you give the oral poliovirus vaccine it can mix with other

01:54:56.180 --> 01:55:03.460
polioviruses in the gut of people and then become active polio again and he's at least

01:55:03.460 --> 01:55:09.300
theoretically aware of the possibility that should you make a de attenuated coronavirus like say

01:55:10.100 --> 01:55:17.300
take out some genes then it would be very easy for that clone that you're using as an attenuated

01:55:17.300 --> 01:55:24.500
virus to attain that gene to acquire that gene from a co-infection and so what he's going to explain

01:55:24.500 --> 01:55:32.580
to you is how these translational regulatory sequences can be altered in such a way to prevent

01:55:32.580 --> 01:55:38.500
recombination with wild type viruses that's his strategy I think it's a pretty cool idea

01:55:39.300 --> 01:55:46.340
it suggests that these self-replicating RNAs are are some kind of a real biological phenomenon

01:55:47.220 --> 01:55:52.980
and it suggests that making infectious clones of them is a really handy way of exploring the

01:55:52.980 --> 01:56:01.140
variation within them that is otherwise un-intractable by laboratory means so here we go again I

01:56:01.140 --> 01:56:07.540
apologize that this is such a fuzzy thing but I will try to help you follow along way for how

01:56:07.540 --> 01:56:12.500
far it gets out of the cell the nuclei caps is the line underneath an intermediate compartment

01:56:12.500 --> 01:56:19.140
in the ruffiardology with a M glycoprotein and the E-protein are expressed and then these the M

01:56:19.140 --> 01:56:24.900
and the E-protein actually drive the assembly of virion which then mature in the Golgi and then

01:56:24.900 --> 01:56:31.940
are released in the cell and secretory vesicles which fuse the last membrane. Now if you knock out

01:56:31.940 --> 01:56:39.780
E-protein expression this pathway is still viable but you reduce virus yields by about two logs

01:56:39.780 --> 01:56:44.900
and if you knock out the M glycoprotein expression or M and E together you don't make any virus

01:56:45.460 --> 01:56:51.620
so M protein will stop without the M protein nothing happens the E-protein will still

01:56:52.660 --> 01:56:57.860
get some viral production I wonder what they mean by that I guess it's just detectable RNA in the

01:56:57.860 --> 01:57:04.100
supernatant. So our approach to de-optimize SARS

01:57:04.100 --> 01:57:14.740
coronavirus the SARS coronavirus genome and to attenuate pathogenesis focused primarily on the

01:57:14.740 --> 01:57:21.460
E and the M glycoprotein genes of about 350 amino acids in total. The strategy we used was to

01:57:21.460 --> 01:57:26.900
progressively increase the number of de-optimized codons so we started with serine to produce a

01:57:26.900 --> 01:57:34.180
D-serve virus serine-loosing R gene to produce the SLR de-optimized strain or a five-set DSLR

01:57:34.180 --> 01:57:39.700
VA strain basically the idea is you create a real stat where you're increasing the number of

01:57:39.700 --> 01:57:44.020
de-optimized residues and turning down expression of critical proteins needed for release.

01:57:45.220 --> 01:57:50.100
So that's an interesting strategy right he's changing the codons

01:57:50.420 --> 01:58:00.180
not changing the codons like to different different amino acids but he's changing the amino acid

01:58:00.180 --> 01:58:08.500
codons so that they're not as they're not what they were in the virus which he assumes is optimized

01:58:09.460 --> 01:58:16.580
which I think is a pretty cool thing because the the assumption for the mRNA

01:58:17.220 --> 01:58:27.780
transfection is that the viral codon selection the the actual codons used by the virus are not

01:58:27.780 --> 01:58:33.380
important and in fact we can change them to whatever we want so that we get more protein

01:58:34.180 --> 01:58:41.300
and we can even chemically alter the RNA with M1 pseudo-uridine alteration so that's fine

01:58:41.300 --> 01:58:49.460
we don't care about that either even though here Ralph Barrick is saying that just by changing

01:58:49.460 --> 01:58:58.260
these synonymous codons for one or two or five amino acids he can progressively attenuate

01:58:59.460 --> 01:59:05.940
viral production he can hamper viral production he can handicap the virus just by changing its

01:59:05.940 --> 01:59:13.620
codons this is pretty antithetical to the idea that Moderna can just change those codons and

01:59:13.620 --> 01:59:20.980
it doesn't matter at all right that's that's pretty impressive this is just a cartoon to show

01:59:20.980 --> 01:59:26.340
amino acid sequence and the wild type virus sequence here so like in the three set at the

01:59:26.340 --> 01:59:32.180
searing residue which was optimal in the case of SARS we just change it to the most

01:59:32.420 --> 01:59:43.460
rare codon if you look at the statistics here in general in the wild type E and M gene they're

01:59:43.460 --> 01:59:50.420
about 39 codons that are de-optimized in the one set this increased to 52 the three set 94 and the

01:59:50.420 --> 01:59:58.900
five set 134 so they're only de-optimizing the codons of the M or the E protein which is even more of a

01:59:58.900 --> 02:00:06.180
subtle modification think about that so look at the ct ctb values that Eckerd just talked about

02:00:07.140 --> 02:00:09.780
by these values here so these are very minimally

02:00:13.140 --> 02:00:20.820
on the negative side of the codon pair usage we also made random controls where we scrambled

02:00:20.820 --> 02:00:25.780
the codon usage much like Eckerd did retaining the wild type genome organization lightless and we

02:00:25.780 --> 02:00:33.300
made three set in the five set this is we made these in a mouse adapted background so they would

02:00:33.300 --> 02:00:40.660
be pathogenic in mice mouse adapted growth is shown here black the blue lines show the searing

02:00:40.660 --> 02:00:47.860
de-optimized viruses which also reach high titers by about 24 hours post-infection if you look at

02:00:47.860 --> 02:00:52.020
the three set mutants however you see about a log log and a half reduction as compared to wild

02:00:52.020 --> 02:00:57.940
type the three set randomized virus and the five set randomized virus in this case i'm showing

02:00:57.940 --> 02:01:03.060
two different plaques and the five set randomized viruses through just about like wild type so just

02:01:03.060 --> 02:01:07.940
like Eckerd had reported if you randomized the sequence it really has very little impact on

02:01:07.940 --> 02:01:15.460
replication but the optimization does affect final yield now in contrast to what Eckerd talked

02:01:15.460 --> 02:01:22.100
about we actually deoptimized in the middle of the downstream ores which would potentially affect

02:01:23.140 --> 02:01:28.340
critical sequences that would affect RNA synthesis we're very careful not to knock out any known

02:01:29.140 --> 02:01:35.780
sequences regulatory sequences that make messenger RNA however both the searing and the SLR mu knocked

02:01:35.780 --> 02:01:40.740
out messenger RNA six expression which you can see right here these actually were the messages

02:01:40.740 --> 02:01:46.340
the genes the messages that encode the gene so this is supposed to be a northern blot where

02:01:46.340 --> 02:01:52.820
the darkness is supposed to show you RNA so this isn't a very clean blot there's an awful lot of

02:01:54.180 --> 02:02:01.540
what is this i mean holy balls you're not getting a clear signal here certainly i mean

02:02:02.100 --> 02:02:12.020
i don't know what i see here but that's not what i would expect to see um he's looking for sub-genomic

02:02:12.020 --> 02:02:18.660
RNAs i guess so this would be the full genome up here i don't know that that's what i would assume

02:02:18.660 --> 02:02:24.580
um it's two hours and i don't think he's going to say very much i'm just going to leave it go but i'm

02:02:24.580 --> 02:02:28.980
going to say very much i'm just going to leave it go but i might put it on a little faster and i

02:02:28.980 --> 02:02:36.420
got to get to bed holy cow it's my birthday today tomorrow so we deoptimized so we actually found

02:02:36.420 --> 02:02:40.020
some evidence of long-range RNA RNA interactions that are regulatory elements that we're trying to

02:02:40.020 --> 02:02:45.620
decipher the five set the optimized trains showed no cpe in culture if you develop primaries to the

02:02:45.620 --> 02:02:49.940
five prime end of the genome and the leader sequence and a primer down here in the message six

02:02:49.940 --> 02:02:53.460
that encodes work six you can actually identify leader containing transcripts which are signatures

02:02:53.540 --> 02:02:57.060
of the messenger RNAs that are made during infected cells in these cultures you can see evidence of

02:02:57.060 --> 02:03:02.260
message seven i'm sorry message six five four and three this is a day one post-transfection

02:03:02.260 --> 02:03:06.020
even by day ten you can still see these transcripts infected cells and they continue at that level

02:03:06.020 --> 02:03:09.540
with about five passages at five-day intervals but you actually never can actually develop

02:03:09.540 --> 02:03:13.140
a virus you can never plaque a virus you can never actually show a virus is there producing cpe

02:03:13.140 --> 02:03:19.140
so that's interesting so now he's looking at how the RNA signal develops over days of passage

02:03:19.940 --> 02:03:24.580
and he he shows how the robustness disappears i think that's kind of cool

02:03:25.460 --> 02:03:32.500
um i don't see a control here though i don't see how the RNA sustains over in days when it's

02:03:32.500 --> 02:03:39.620
not attenuated that would be a nice control to have here right um anyway i think this is where

02:03:39.620 --> 02:03:45.060
he's going to end it he's probably just going to close he had deoptimization he's going to show

02:03:45.060 --> 02:03:50.980
these three different ways that they did it and the sites that they changed and then he shows

02:03:50.980 --> 02:03:58.260
pathogenesis of these things in the mice he made a mouse adapted version of it for aged mice um

02:03:58.260 --> 02:04:03.220
it's an it's an it's an oh these are all papers that we've looked at before they're all relevant for

02:04:03.860 --> 02:04:12.900
the clone discussion because the clone discussion puts us in um the context of the pandemic and

02:04:12.980 --> 02:04:19.300
trying to explain what's happening here i want to just leave you with the slide that i put up

02:04:19.300 --> 02:04:30.260
earlier um with regard to alina chan um and kevin mccurnan's objection that uh maybe there should be

02:04:30.260 --> 02:04:37.940
some some so ask yourself why he's making six-hour scooby-doo videos instead of downloading data

02:04:38.420 --> 02:04:43.700
from ncbi and find a signal for his hypothesis in real data if the clones offer some advantage

02:04:43.700 --> 02:04:51.620
for dr evil surely there is evidence of reduced mutation rates to prove this and i would humbly

02:04:51.620 --> 02:04:59.140
submit that actually alina chan and employee of the broad institute for whom your uh old boss is

02:04:59.140 --> 02:05:04.980
now the director for quite some time um has data from the beginning of the pandemic that seems to

02:05:04.980 --> 02:05:15.300
show a much lower mutation rate in the sars 2 sequences than those found during a similar time

02:05:15.300 --> 02:05:22.980
frame in the sars pandemic which might be the reduced mutation rate you were looking for um

02:05:22.980 --> 02:05:28.340
we have a long ways to go we got a lot of stuff to talk about but i'm not really worried the reason

02:05:28.340 --> 02:05:32.500
why i'm not worried is because people have lose in their minds and when people lose their minds

02:05:32.580 --> 02:05:37.380
it usually means that you're right over the target have never seen so many people lose their

02:05:37.380 --> 02:05:42.900
mind at once and i've never seen so many people do it that weren't doing it together before

02:05:43.700 --> 02:05:49.460
um people are coming together and retweeting and talking to one another and patting each other

02:05:49.460 --> 02:05:55.700
on the back that heretofore have never really interacted in that way before and that's pretty

02:05:56.020 --> 02:06:03.780
special not only that but it's brought together a few of these dream team members and uh i think

02:06:04.340 --> 02:06:14.500
the spontaneous sort of confluence of efforts are going to become obvious what everybody's

02:06:14.500 --> 02:06:19.700
arguing about is we all got to come together because because because i'm making exactly the

02:06:19.700 --> 02:06:24.740
opposite argument i don't need to come together with anybody because if we are fighting for the

02:06:24.740 --> 02:06:31.140
same things then our our efforts will buy necessity and buy natural

02:06:32.660 --> 02:06:38.100
by natural order where they will they will become a confluence of effort toward the same goals

02:06:38.740 --> 02:06:45.540
my goal is to have our children realize that the vaccine schedule in america is a criminal

02:06:45.540 --> 02:06:52.420
enterprise my goal is to have our children realize that virology especially RNA virology

02:06:52.420 --> 02:06:58.180
is wholly dependent on cloning technology and in fact a pandemic could not have occurred in

02:06:58.180 --> 02:07:02.820
the way that they said it is no matter how much they insist that the sequences are here and not

02:07:02.820 --> 02:07:09.220
there or that they're real or they're not they are lying about the fidelity and that's all that i

02:07:09.220 --> 02:07:15.940
can say and and all this this insistence is not good enough they need to start teaching because

02:07:15.940 --> 02:07:21.780
that's what we're doing that's what we've been doing for three and a half years no one else is doing it

02:07:21.780 --> 02:07:26.100
just us just here and so if you like what we're doing you like what we've done

02:07:27.060 --> 02:07:32.180
please support our work go to get your own biological and subscribe share the work if you can

02:07:33.700 --> 02:07:41.940
we are gaining momentum tomorrow is my birthday but i will be on i'm going to try and be on as

02:07:41.940 --> 02:07:46.500
often as literally as often as possible i don't know if it's going to be every day or not

02:07:47.380 --> 02:07:53.940
um but i really am targeting every day but i also have other things that i'm trying to get

02:07:53.940 --> 02:07:59.060
regular with including the sub stack i don't think i'm going to make a sub stack and transcript of

02:07:59.060 --> 02:08:03.540
this one but i want to do it for a lot of them and so i don't know if you've noticed it or not but

02:08:03.540 --> 02:08:11.140
there is a sub stack now the sub stack has videos that are on vimeo um those videos i've added

02:08:11.700 --> 02:08:19.780
uh i've added subtitles to and so that makes them also kind of um better to watch you can also

02:08:19.780 --> 02:08:23.860
share them a little bit better the sub stack maybe is a better way to share them because people can

02:08:23.860 --> 02:08:32.340
scan the the the transcript as well and so wolf gang wodocks interview is up there um my presentation

02:08:32.340 --> 02:08:38.660
to the uk doctors is up there um and uh i'm just going to keep trying to do it every time i get a

02:08:38.660 --> 02:08:43.860
stream that i think is good enough or is worthy of it um we're going to do a sub stack on it i

02:08:43.860 --> 02:08:49.460
don't think i'm going to do this one because tomorrow i'm going to try and do if all goes well

02:08:49.460 --> 02:08:55.460
i'm going to try and do a pretty decent show tomorrow um but it's a birthday and i've got two

02:08:55.460 --> 02:08:59.460
basketball games so i don't know there's going to be a show and it's don't know if it's going to be

02:09:00.660 --> 02:09:06.100
slam-bam crazy show or if it's just going to be an average show like this one was but anyway

02:09:06.820 --> 02:09:12.180
we've got the momentum going again or at least i got it going tonight and so hopefully i can get

02:09:12.180 --> 02:09:19.540
some sleep and uh get it going tomorrow again and uh we'll get this this pace up to speed thank you

02:09:19.540 --> 02:09:35.460
very much for joining me and i will definitely see you again tomorrow

02:09:49.540 --> 02:10:16.020
i have no responsibility for the current pandemic

