Tired of Waiting - Hiring CRISPR scientist now

[mcmich]

Hi

I know there was a thread about hiring a scientist to run CRISPR experiments on those of us that are brave enough to try it. A few people on this board even seemed qualified to try it themselves.

Anyway, is there any company, whether here in the US or overseas that these experiments could be contracted through? Be nice to get a cure now and not wait 10 to 20 years!!!!

[impugn]

I've thought about this before. I have no idea why there are restrictions on things of this matter. If you accept the risks and your doctor accepts the possibility that he could harm you and is comfortable with the chance of that happening, there should be no reason we legally can't. There are millions of people who suffer bad enough from herpes simplex that they are willing to die. If they are willing to die, they would absolutely be willing to be a guinea pig if they have a chance of a cure or functional cure. There's no reason we can't choose our fate. 

[dont quit!17]

2 hours ago, mcmich said:

Hi

I know there was a thread about hiring a scientist to run CRISPR experiments on those of us that are brave enough to try it. A few people on this board even seemed qualified to try it themselves.

Anyway, is there any company, whether here in the US or overseas that these experiments could be contracted through? Be nice to get a cure now and not wait 10 to 20 years!!!!

CHINA. If there ever is a promising CRISPR therapy, Im sure you can find some lab and or scientist that can replicate the therapy as well as administer the therapy itself. It would cost a pretty penny but if effective it would truly be amazing. China was First Nation to actually run trials with CRISPR.

http://fortune.com/2016/11/15/first-crispr-trial-humans-china/

Maybe in a few years we can go there for HSV therapy. Maybe after a promising phase 1 from any US ran clinical trial. I may have to brush up on Mandarin. 

[Sanguine108]

Yeah, how far-fetched would it be for a group of people outside of China to fund a lab space with some workers/scientists/techs in China and have someone who seems passionate and competent enough to fly over and/or live over there/supervising and whatnot. (cough OFMDH and/or StayingUpbeat).

If it works then we gotta take a trip to China to the treatment.

[Seeker1960]

I think this idea sounds good but way premature. I think Crisper Cas9 shows promise but not develop anywhere near enough to try on humans. There could be some good developments soon as animal trials are starting soon. This would surely kill you if it wasn't 100% correct.

Theravax is riskybalso but at least it is based on familiar technology and has gone through animal testing and human tests.

Lets see after another year

[fugazi1]

 

[dont quit!17]

1 hour ago, Seeker1960 said:

I think this idea sounds good but way premature. I think Crisper Cas9 shows promise but not develop anywhere near enough to try on humans. There could be some good developments soon as animal trials are starting soon. This would surely kill you if it wasn't 100% correct.

Theravax is riskybalso but at least it is based on familiar technology and has gone through animal testing and human tests.

Lets see after another year

Yeah, it would be nothing immediate but 5 years worth of research and I believe some solid techniques should come to light in regards to CRISPR and very possibly the actual strategy to cure the virus. Who knows, maybe we can take the Excision strategy, take it to China, get cured and enjoy a greater quality of life. We should ALL get together in order to see IF something like this could happen. 

Unfortunately, I don't think Theravax is for everyone but I hope Im proven wrong. One years worth of research should tell us a lot. Genocea is planning to run trials with vaccine + antiviral combo. HSV-29 phase 1 trials should be released as well. And it would be interesting to see what Admedus does with their product. 

[jreemi]

Crispr has already been tried in humans (china) and the one for hsv so far looks safe (only edited the target gene, nothing else affected)

[LillianPanos]

Where did u get this info

11 hours ago, jreemi said:

Crispr has already been tried in humans (china) and the one for hsv so far looks safe (only edited the target gene, nothing else affected)

 

[Atrapasueños]

Esta nota esta en español pero enfatizan que China esta a un paso más adelante que Europa y Estados Unidos en cuestión de usar crispr 

Ahora hay una noticia donde hay gente que está creando empresas para curar sus propias enfermedades usando crispr

 

 

This note is in Spanish but they emphasize that China is a step further than Europe and the United States in question of using crispr

Now there is a story where people are creating companies to cure their own diseases using crispr

http://www.abc.es/ciencia/abci-carrera-mundial-para-editar-genes-y-curar-enfermedades-201611162250_noticia.html

 

[Atrapasueños]

Se menciona que el tratamiento de crispr si todo va bien sería dentro de 5 años pero no se afirma que sea una cura en ese lapso de tiempo sólo se indica que es para el que tiene el virus

 

It is mentioned that the treatment of crispr if everything goes well it would be in 5 years but it is not affirmed that it is a cure in that time span only indicates that it is for the one who has the virus

[Atrapasueños]

https://www.technologyreview.es/biomedicina/53323/una-asociacion-de-pacientes-invierte-millones/

asociación de pacientes invierte para curar su propia enfermedad distrofia muscular

Association of Patients Invests to Cure Their Own Disease Muscular Dystrophy

[Renegadow]

On 4/1/2017 at 1:18 AM, Sanguine108 said:

Yeah, how far-fetched would it be for a group of people outside of China to fund a lab space with some workers/scientists/techs in China and have someone who seems passionate and competent enough to fly over and/or live over there/supervising and whatnot. (cough OFMDH and/or StayingUpbeat).

If it works then we gotta take a trip to China to the treatment.

Someone passionate enough to formulate a real project and materialize the idea , and people from HC can donate and support.

There's a lot of talk n this forum with zero action. We are a bunch of people, we  all have the same problem, we need to get together ,stop crying/complaining/spamming all over the place and start thinking and acting rationally,  Because that's the only way out of this.

And I'll give you a hand with @'s

@OhFuckMyDickHurts

@StayingUpbeat

[fugazi1]

I agree, there is so much potential here to rally together and make a bit of noise. I wish I knew where to start though. 

[StayingUpbeat]

I am the last person who wants to rain on anyone's parade.  As the name implies I am positive about the future and what it holds.  Something that everyone needs to accept and get past to see that positive future, is that we will almost certainly have herpes (though with likely fewer symptoms) for the rest of our lives.  A sterilizing cure, without getting into the science, is simply not how CRISPR works. 

With that out of the way there are three basic problems with a grass roots effort to cure herpes using a manufacturing driven country like China.

1. There is still a lot of fundamental research necessary that China is not well positioned for
2. Herpes has the numbers that would imply a lot of money available but seems only be able to generate hard cash on the order of thousands of dollars.
3. China is great at making things but not so much at developing new things

When espousing the idea of a grass roots funded campaign to cure herpes, in any promising technology: "all of this has happened before and will happen again."

• The herpes cure coalition  and the Pepsi Refresh everything campaign. (circa 2008)
  • Raised $50K for Dr. Cullen and Bloom's Pre-CRISPR gene editing concept
• Broken_14's quest DRACO and the Killing Sickness Campaign (circa 2015)
  • Raised ~$100K for Dr. Rider's DRACO broad spectrum antiviral concept

The idea would border on plausible if there was something relatively close technologically.  For instance it's not hard to make DRACO.  I synthesized it as an independent projects for one of my biotech classes.  Since I wanted a control to compare it with I had a company synthesize a few milligrams of it for about $1K. 

Having the CRISPR guide sequence, a viral vector, and willing human guinea pigs is <1% of the equation.  Practically every disease with a DNA basis has those ingredients.  The problem is that making it work safely in people is very, very, hard.  Even with an infinite budget I am not close to the level of smart it would require. 

Now will CRISPR (or some evolved form of it) one day be able to change any DNA in all your cells?  Maybe.  In the type of future where you can go to your geneticist and have your hair or eye color changed it seems perfectly plausible that he/she can cleave all the HSV DNA out too.  I simply don't see that, with the state of technology today, happening in a timeframe where I'm not long gone. 

I do, however, think it's likely we'll see something that can knock a big chunk of the viral reservoir out (greatly reducing symptoms) with prophylactic vaccines keeping the people we care about safe.  Technology seems to be marching that way but its still going to be quite a while and there's very little a few thousand (or even a few million) dollars is going to do to accelerate that.

[Atrapasueños]

The pepsi contest that won Dr bloom in collaboration with Dr Cullen did not use crispr

And draco was simply not promising for herpes since its functionality was to kill infected cells but not latent viruses in neurons

I think the best option is that people pressure governments with campaigns for a cure, as they do with hiv and cancer if you do not do that, nothing will speed up the process

[Cas9]

1 hour ago, StayingUpbeat said:

I am the last person who wants to rain on anyone's parade.  As the name implies I am positive about the future and what it holds.  Something that everyone needs to accept and get past to see that positive future, is that we will almost certainly have herpes (though with likely fewer symptoms) for the rest of our lives.  A sterilizing cure, without getting into the science, is simply not how CRISPR works. 

With that out of the way there are three basic problems with a grass roots effort to cure herpes using a manufacturing driven country like China.

1. There is still a lot of fundamental research necessary that China is not well positioned for
2. Herpes has the numbers that would imply a lot of money available but seems only be able to generate hard cash on the order of thousands of dollars.
3. China is great at making things but not so much at developing new things

When espousing the idea of a grass roots funded campaign to cure herpes, in any promising technology: "all of this has happened before and will happen again."

• The herpes cure coalition  and the Pepsi Refresh everything campaign. (circa 2008)
  • Raised $50K for Dr. Cullen and Bloom's Pre-CRISPR gene editing concept
• Broken_14's quest DRACO and the Killing Sickness Campaign (circa 2015)
  • Raised ~$100K for Dr. Rider's DRACO broad spectrum antiviral concept

The idea would border on plausible if there was something relatively close technologically.  For instance it's not hard to make DRACO.  I synthesized it as an independent projects for one of my biotech classes.  Since I wanted a control to compare it with I had a company synthesize a few milligrams of it for about $1K. 

Having the CRISPR guide sequence, a viral vector, and willing human guinea pigs is <1% of the equation.  Practically every disease with a DNA basis has those ingredients.  The problem is that making it work safely in people is very, very, hard.  Even with an infinite budget I am not close to the level of smart it would require. 

Now will CRISPR (or some evolved form of it) one day be able to change any DNA in all your cells?  Maybe.  In the type of future where you can go to your geneticist and have your hair or eye color changed it seems perfectly plausible that he/she can cleave all the HSV DNA out too.  I simply don't see that, with the state of technology today, happening in a timeframe where I'm not long gone. 

I do, however, think it's likely we'll see something that can knock a big chunk of the viral reservoir out (greatly reducing symptoms) with prophylactic vaccines keeping the people we care about safe.  Technology seems to be marching that way but its still going to be quite a while and there's very little a few thousand (or even a few million) dollars is going to do to accelerate that.

"Something everyone needs to accept......is that we will almost certainly have herpes (though with likely fewer symptoms) for the rest of our lives".
Thank you professor.
You know I wasn't sure what the future holds but now that you've told us what's going to happen I guess you've answered all our questions about the future. Have you contacted all the professors of all the research labs and told them to stop working on a sterilizing cure? Please do because we wouldn't want them to be wasting there time.

As a side note, since you said "that's not how CRISPR works", then explain how it works.

And a sterilizing cure, whether it comes or not, is irrelevant. There are probably viruses in you right now that you never in even heard of. It's a non issue because they cause no disease. Therefore, if herpes can be rendered to a status where no virus replicates, or at least doesn't replicate at the skin surface, then it falls in the category as all the other insignificant viruses you may have. No sores, no symptoms.

[Atrapasueños]

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4928872/ 

Este artículo es muy completo y científico sobre como combaten los herpesvirus vs crispr

 

This article is very complete and scientific about how they fight herpesvirus vs crispr

[OFMDH]

1 hour ago, StayingUpbeat said:

........The idea would border on plausible if there was something relatively close technologically.......

Having the CRISPR guide sequence, a viral vector, and willing human guinea pigs is <1% of the equation............

....the problem is that making it work safely in people is very, very, hard.......

A problem I found with China is they still buy all, or mostly all of their supplies and equipment from the USA to run an experiment so the savings are not that great when you run the figures. Further it takes 2-3 weeks to get anything shipped there or visa versa and they don't publish research in English. I find it quite hard to decipher their research publications on HSV at times. Further while they are playing catch up in a lot of areas and working on Cas9 I still don't think they're going to share everything with the world, or private lab there what they find out anyways. 

Safety is definitely a huge concern and one of the prime reasons the Chinese human trials using Cas9 was done in such a way to verify the changes outside the body in the cell and then re-injected the edited cells back in. Safety... its a moot issue as what really holds back research and innovation for HSV to even ask that question is simply the business case for something better than $5/monthly generic Valtrex. Looking at Genocea's S.E.C. filings they go over this and estimate a cap in market size and price; it's not pretty. That being said there are patent holders already testing Cas9 against HSV but I doubt they'll try it with a vector to go after the latent DNA in the body.

Further there are things that can be tried out to perfect the use of Cas9 in a vector against HSV, heck a lot of different things can be tried as well besides Cas9 in a vector or even with Cas9, and even new potential antivirals. But for those wanting Cas9 [and the sgRNA's] in a viral vector.... I have moved already on this and I'd really want to use and verify it in vivo by a lab and looking at what happens to the latent DNA over the long term [over 6 months]. Note, kinetics of Cas9 are much slower on latent DNA, but contrary to other experiments it still can get at it. It is possible to inject a person, but it would not be verified as either safe or effective. Otherwise the cheapest way to do all this is just get a NIH grant... but I'm not actually killing mice and living in a lab. I'd rather set it up and hire someone to do the grunt work. It doesn't mean it will come to market though in the USA. 

 

 

[Cas9]

6 hours ago, Atrapasueños said:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4928872/ 

Este artículo es muy completo y científico sobre como combaten los herpesvirus vs crispr

 

This article is very complete and scientific about how they fight herpesvirus vs crispr

I pretty much know how it works. So I wasn't requesting that StayingUpbeat explain for me if this article was for me; I wanted StayingUpbeat to explain his understanding to see how accurate he is. In other words, I was testing him.
That said, he may not be accurate OR he may lend some information that I don't know.

[OFMDH]

12 hours ago, Atrapasueños said:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4928872/ 

Este artículo es muy completo y científico sobre como combaten los herpesvirus vs crispr

 

This article is very complete and scientific about how they fight herpesvirus vs crispr

This isn't in vivo though. The problem is getting Cas9+sgRNA's to the cells with latent DNA in them and then the efficiency of the snipping of DNA and time period necessary to do that and whether the vector can last enough to do it. 

Contrary to statements that the virus has to replicating to be accessible to be edited, over the long term Cas9 does indeed go into heterochromatic DNA (aka latent viral DNA).  The kinetics are just a lot slower. Thus the next question is what happens after the snipping when the cell repairs it and whether if viral DNA is replicating what the the repair efficiency is. There will always be a combination of desired edits to stop replication and some that don't work. Thus multiple sgRNA targets are needed. The more targets though, the less a vector will express over time. Then it becomes a balancing between the time / kinetics of CRISPR in latent viral DNA with sgRNA's and whether the vector will express all of that long enough to get at it with the desired results (assuming you can get 99.99% transfection rate to get at every cell with that viral DNA). The highest transfection rates right now are with HSV1 vectors. Thus its the best candidate to use as a vector. 

Although an issue has also been the use of viral promoters to drive the CRISPR (Cas9/Cpf1, etc.) and the sgRNA's expression within the cell, there are actually human proteins that can be used instead as promoters to drive expression -- they all have differing degrees of efficiency/longevity. The other issue is leaving antibiotic selection genes in the vectors from the plasmid generation. FDA recommends removing those for safety reasons, if it can be done. Otherwise they'll allow for testing in humans only two such different genes in a vector. 

[jreemi]

18 hours ago, StayingUpbeat said:

I am the last person who wants to rain on anyone's parade.  As the name implies I am positive about the future and what it holds.  Something that everyone needs to accept and get past to see that positive future, is that we will almost certainly have herpes (though with likely fewer symptoms) for the rest of our lives.  A sterilizing cure, without getting into the science, is simply not how CRISPR works. 

Well for one, I'm not convinced that you understand how CRISPR works.

I will have chicken pox for the rest of my life, but it is not a factor. One can carry a virus but not have disease. The earliest studies of CRISPR on HSV has shown extremely potent inhibition of the virus. The current issue at hand is reaching the latent virus which lies in non-replicating/closed off neurons. That can and will be overcome in the near future. You dont know what will happen tomorrow. They could cleave the DNA in vitro next year. To say that we'll die with it, well, is misleading at best and irresponsible at worst. I haven't heard the most pessimist of scientists proclaim it won't have a fix within a lifetime. A full blown cure, which you seem to forget, is not necessarily needed to make the virus irrelevant.

As far as safety is concerned, this system is also promising:

"We found our Cas9/gRNA system lacks adverse effects upon cell viability and processes including including the cell cycle, does not induce apoptosis, and does not jeopardize cell viability. To achieve this level of safety, avoid effects on human translated genomic sites and exclude any transcription DNA motif, we employed bioinformatic screening based on the strictest 12 bp plus PAM target selection criteria. Our most effective gRNA (2A) was 30nt in length plus NGG. It showed no evidence for InDel mutation in any of a series of five representative off-target host genes identified by bioinformatic screening using shorter (12 nt) seed sequences corresponding to target A of ICP0 exon II. Our results, also show that the CRISPR/Cas9 system can be refined and used for protecting cells against HSV-1 infection"

-http://media.wix.com/ugd/54ec3c_17b495ebfee84a7eaaa4fd51b9634db7.pdf

There are several other studies I've read, in detail, and the prevailing theme is that specificity with regards to targeting can easily be achieved with Cas9 and more importantly that other cells are not affected. There is work to do, but safety won't be as big of a mountain to climb as you make it seem.

 

[OFMDH]

3 hours ago, jreemi said:

Well for one, I'm not convinced that you understand how CRISPR works. 

..... The current issue at hand is reaching the latent virus which lies in non-replicating/closed off neurons. That can and will be overcome in the near future. You dont know what will happen tomorrow. They could cleave the DNA in vitro next year.

A little civility is nice once in awhile.

I just said in the prior post Cas9 proteins do indeed make forays into latent DNA in vivo, albeit the kinetics are a lot slower. Even the FHCRC showed with HE you did not need replicating viral DNA to insert mutations - while they had a low # of mutations found, they did not go longer term. If I do the math I can probably figure it all out that indeed eventually you can get all the viral DNA. Otherwise the in vitro cell dishes experiments with HSV and Cas9 don't pick this fact up because it takes a lot longer to happen.

Quote

As far as safety is concerned, this system is also promising.......... There are several other studies I've read, in detail, and the prevailing theme is that specificity with regards to targeting can easily be achieved with Cas9 and more importantly that other cells are not affected. There is work to do, but safety won't be as big of a mountain to climb as you make it seem.

The safety issue is not the off-target concerns. Cas9 for instance "likes" or rather works best with high GC content DNA that is luckily different enough from human DNA that the sgRNA targets are usually quite far off enough that it would require more than just a several major mistakes in the sgRNA itself to start going after human DNA. It's more likely the expression of sgRNA's would stop before that happens. 

Rather one safety issue is inserting viral promoters and antibiotic resistant genes into vectors to go after the latent HSV DNA in our bodies and all of it together overworking the cell's machinery eventually. Otherwise as Cas9 is a foreign protein it will just break down in the cell through the regular enzymes there. The vector just has to keep expressing it (make more Cas9+sgRNA's) to keep going after the HSV DNA but not so long that it wears out or clogs the cell's own machinery. Thus there is even more complexity in developing down the road maybe a means to repress expression. However I personally think if the vector wears off fast enough you can avoid the eventually clogging effect and finding a means to suppress the expression. 

One potential unknown whether or not the Cas9 protein in the human cell in the nucleus will invoke an immune response from the human. As far as I know it does not. Otherwise "CRISPR" systems are still advancing.

The future is wonderful. 

 

[OFMDH]

On 4/3/2017 at 9:01 PM, StayingUpbeat said:

.....As the name implies I am positive about the future and what it holds............... we will almost certainly have herpes...........for the rest of our lives.  A sterilizing cure, without getting into the science, is simply not how CRISPR works. 

Long term studies show that using antivirals every year there is a % drop of those who don't have OBs, such that by year 5 still over 20% will not have had an OB while on pills.

In regards to having it.... 

If got Cas9 to all the neurons with the required sgRNA's in a vector I would have herpes DNA technically in neurons but I may not have herpes disease on my genitals/skin. That's because replication would be disabled. This is not a foreign concept, again on antivirals a large percent of the infected cells with the virus eventually chain terminates its replication and fragments just build up. These fragments eventually get broken down and recycled by the cell. Thus the virus is not able to copy all of its DNA and fails to build new virus. A reason its not 100% at stopping all shedding is that antivirals are still competing with other chemicals in the cell, thus while its incorporated and messes up the viral replication sometimes its not incorporated.  

Maybe down the line not only can one silence or knockout the viral DNA replication, but cause the neurons to also eject the defunct viral DNA from the nucleus and signal that it should be broken down. That I think will be difficult. Otherwise people get too hung up on the idea of a sterilizing cure as opposed to a functional cure. If I were to disable replication of all viral DNA and never have another OB then it doesn't matter if the cell has defunct viral DNA in it. It's never going to cause any visible disease or bother me again. It's now just a skeleton in the closet I can forget about.

Out of sight, out of mind.

[valleynovascotia]

3 hours ago, OhFuckMyDickHurts said:

Long term studies show that using antivirals every year there is a % drop of those who don't have OBs, such that by year 5 still over 20% will not have had an OB while on pills.

In regards to having it.... 

If got Cas9 to all the neurons with the required sgRNA's in a vector I would have herpes DNA technically in neurons but I may not have herpes disease on my genitals/skin. That's because replication would be disabled. This is not a foreign concept, again on antivirals a large percent of the infected cells with the virus eventually chain terminates its replication and fragments just build up. These fragments eventually get broken down and recycled by the cell. Thus the virus is not able to copy all of its DNA and fails to build new virus. A reason its not 100% at stopping all shedding is that antivirals are still competing with other chemicals in the cell, thus while its incorporated and messes up the viral replication sometimes its not incorporated.  

Maybe down the line not only can one silence or knockout the viral DNA replication, but cause the neurons to also eject the defunct viral DNA from the nucleus and signal that it should be broken down. That I think will be difficult. Otherwise people get too hung up on the idea of a sterilizing cure as opposed to a functional cure. If I were to disable replication of all viral DNA and never have another OB then it doesn't matter if the cell has defunct viral DNA in it. It's never going to cause any visible disease or bother me again. It's now just a skeleton in the closet I can forget about.

Out of sight, out of mind.

Would this make us none contagious as well?