PLAUSIBLE CURE - CRISPR/CAS9 GENE EDITING

flynn

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Feb 28, 2018
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#1
CRISPR

It may seem a bit crazy, but I really believe if we are ever to find a working, effective treatment/cure, it will likely come from Gene therapy. Luckily for us, the CRISPR gene editing system has arrived and offers the potential to fix this damage.

If a theory such as the 5AR theory of PAS is true (or PAS relates to changes in another type of gene), I think CRISPR may be an ideal treatment candidate. CRISPR enables cheap, fast, precise and efficient gene editing. This technology is very new but is expected to make medical therapies which target and work at the genetic level, a reality.

Why CRISPR? It appears that Accutane may have altered the expression of specific genes, resulting in a persistent downregulation which continues after ceasing treatment. A possible candidate are the genes encoding the 5 alpha reductase (5AR) enzyme, specifically 5AR type 1. See here for more info - https://pasforum.info/threads/theor...y-seems-to-be-the-most-likely-cause-of-pas.9/

CRISPR is not just capable of inserting genes or removing/silencing them. It can also enhance the expression of a desired gene in a process known as CRISPR based gene activation. Given this, CRISPR could be used to significantly increase the expression of the 5AR genes, resulting in an increase in 5AR expression/activity and hopefully alleviation of some of the persisting side effects of PAS.

This may seem a bit like science fiction, but it's already possible to purchase 5AR1 and 5AR2 CRISPR gene activation plasmids online for both human and mouse cells (although these won't work for neurons yet as explained later) - https://www.scbt.com/scbt/browse/5-alpha-reductase-Antibodies/_/N-egnv73#CRISPRs2

Moreover CRISPR technologies such as these, may also be very useful tools in studying the development of PAS in animal models. For example, the brains of mice could treated CRISPR products which reduce the expression of the 5AR1 gene by a factor, which mimics the effect of Accutane. To note, a study in human skin found that Accutane treatment resulted in a 2.83 fold reduction in 5AR1 expression. Researchers could then study how this effects mice behaviour/activity/sexual responses etc. These models could then be used to test the effect of using a CRISPR gene activator, to see if mice regain their normal behaviour/activity/sexual function. This would help elucidate the cause of the condition and provide impetus for some kind of clinical trial.

Where and how do we target the brain?

The most difficult thing about using CRISPR will be getting a delivery method which targets the brain as its very difficult to get drugs/molecules/therapies through the blood brain barrier (BBB). It will also need to reach the right areas of the brain which have been affected. This will no doubt be a challenge. But I envision many researchers are already developing CRISPR delivery methods for the treatment of a range of neurological disorders which could be copied/used.

In regards to the brain areas to target, at this point, its a guessing game. But according to the following study in mice, the conversion of testosterone to DHT and thus 5AR activity is very high in the pituitary gland and hypothalamus. Other areas with less activity are the cerebral cortex and amygdala -
https://app.dimensions.ai/details/publication/pub.1042917474

Given the prevalence of retinoid receptors in the brain and the ability of Accutane metabolites such as retinoid acid to permeate the brain, its possible that gene expression in all these areas has been altered. Thus efforts could either try a general approach which targets the whole brain or a specific approach which targets individual brain regions. So how could researchers get the therapy inside the brain? One method may be to inject the gene therapy directly into the brain to bypass the Blood Brain Barrier, its called intracerebroventricular injection. It would require finding a doctor willing to do it, but this has and can be done. One method of intracerebroventricular administration is called Ommaya reservoir. If its a gene therapy, this would be viable as a person may only need to do it once or twice for lasting effect -
https://www.ncbi.nlm.nih.gov/pubmed/19558257

Cutting edge medical researchers are experimenting with this kind of technology, using MRI guidance to inject a gene therapy directly into the brain to treat brain cancer. Showing it can be done, I imagine this kind of technology will become a standard in the future as it removes the complications surrounding the BBB and allows highly targeted delivery -

Another hope is that new gene therapy delivery systems will be able to deliver the CRISPR components across the BBB and into the brain. Fortunately teams are working on developing viral vectors to act as delivery systems for gene editing tools such as CRISPR, which specifically target the brain - http://www.caltech.edu/news/novel-viral-vectors-deliver-useful-cargo-neurons-throughout-brain-and-body-78785

Despite this, there is another challenge which is that currently the most effective version of CRISPR only works on dividing cells. So it wouldn't be very effective on neurons as for the most part, these cells are non-dividing. Therefore a treatment option would need to use an adapted version of CRISPR. Fortunately, methods such as these are already being developed, given the huge clinical potential of gene therapies for neurological disease, I envision in the future, there will be a range of highly advanced techniques for the use of CRISPR in the brain -
https://www.fiercebiotech.com/resea...r-neurons-could-boost-research-brain-diseases

Who would develop it/How would we be able to use it?

So how or who would develop a treatment such as this? How could we ever get access to this technology. At present, it seems very unlikely that pharma companies will take much notice or interest in treating these conditions. One option may be bio-hacking/self experimentation. Now I don't condone this and recognise that currently most attempts to bio-hack are both very stupid and likely to be mostly ineffective. But bio-hacking/self experimentation is both possible and is already taking place, in some cases with experimental gene therapies. Over time, the cost required to use gene editing tools is likely to continue to decline whilst the efficiency and specificity (safety) is likely to improve.

Hypothetically, if the community managed to raise sufficient funds, a CRISPR based system such as the ones linked above which work in neurons could be purchased and tested on lab rats and mice in a research project. Provided an appropriate delivery system was available. A brave individual from the community, could then try small dosages of it, and incrementally build up the dosage to see if there is any beneficial effect. It would likely involve finding a doctor who worked in a country with less regulation. For example, Biotech CEO Liz Parrish had to travel to Colombia to have an experimental gene therapy injected into her as American doctors weren't allowed to by law (https://www.theguardian.com/science/2016/jul/24/elizabeth-parrish-gene-therapy-ageing). This just shows that it is possible, in the future, surgeries which target therapies directly to the brain may be very prevalent and cheap. Thus just as Liz Parrish traveled to Colombia to have a gene therapy injected in her body, it may be possible to have one injected into the brain.

Now I admit, this is all a little far fetched and wouldn't be possible right now. But this field of research is advancing very rapidly, with enormous amounts of innovation. Soon these CRISPR technologies will be cheaper, safer and even more targeted than today. There are already numerous individual who are experimenting on themselves with gene therapies, I imagine this will continue in the future. Here are some examples:

https://www.technologyreview.com/s/...lf-with-a-diy-herpes-treatment-live-on-stage/

https://www.ft.com/content/ed204b7a-314d-11e8-b5bf-23cb17fd1498

https://www.nextbigfuture.com/2017/11/personal-crispr-genetic-experimentation.html

- Flynn
 
Last edited:

Lost

New member
Mar 10, 2018
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#2
The problem would be generating enough interest in the researchers who will perform these. Pas victims have low visibility because everybody thinks its in our brains, nice finding though brother
 
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flynn

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Feb 28, 2018
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#3
The problem would be generating enough interest in the researchers who will perform these. Pas victims have low visibility because everybody thinks its in our brains, nice finding though brother
Yes thats true. But there are several things to note. Firstly, over the coming years I imagine PAS will become recognised in the same way as PFS has. There is already growing awareness thanks for organisations such as RxISK and as long as people keep reporting side effects. There needs to be. But even if it doesn't.

Post Finasteride Syndrome (PFS) would require pretty much the exact same treatment/technique. PFS is already recognised as a condition. So if a treatment was developed for PFS, either PAS people could use it directly or they could tweek it slightly and then used it.

Lastly, its not out of the question that you could get access to something like this through self experimentation or private labs. As this technology develops, the ease of using it will only improve. In several years time, you may be able to just buy custom CRISPR gene edit/activators which you could self-administer to see what happens (high risk but people are already trying it out).
 
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tom.math

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Mar 10, 2018
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#4
Yes thats true. But there are several things to note. Firstly, over the coming years I imagine PAS will become recognised in the same way as PFS has. There is already growing awareness thanks for organisations such as RxISK and as long as people keep reporting side effects. There needs to be. But even if it doesn't.

Post Finasteride Syndrome (PFS) would require pretty much the exact same treatment/technique. PFS is already recognised as a condition. So if a treatment was developed for PFS, either PAS people could use it directly or they could tweek it slightly and then used it.

Lastly, its not out of the question that you could get access to something like this through self experimentation or private labs. As this technology develops, the ease of using it will only improve. In several years time, you may be able to just buy custom CRISPR gene edit/activators which you could self-administer to see what happens (high risk but people are already trying it out).

will need a lot of awareness and maybe organisation like PFS to get mainstream
 
Likes: flynn
Mar 27, 2018
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#5
Crazy or not...seems like the most positive/likely thing I've come across on my trawls of the web. Impressed, Flynn, by your optimism. I'm not a scientist: are we saying then, that the epigenetic changes from Accutane are reversible in theory?

Several years for a therapy though, even if they can find one...it's been 31 for me, not sure how many more I can take!
 
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flynn

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Feb 28, 2018
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#7
Crazy or not...seems like the most positive/likely thing I've come across on my trawls of the web. Impressed, Flynn, by your optimism. I'm not a scientist: are we saying then, that the epigenetic changes from Accutane are reversible in theory?

Several years for a therapy though, even if they can find one...it's been 31 for me, not sure how many more I can take!
Thanks for the response. It's the main reason I made this forum, as so much of the talk regarding this syndrome is merely people complaining and wishing it had never happened. People also make pretty big assertions without really thinking about the causes without providing much evidence or thought about the likelihood of that theory. The reason I made the theories of PAS, is to present each theory and weigh it up. I honestly think some of the darker theories as to how PAS arises relating to schizophrenia or brain damage, don't hold as much weight as people like to think. And it gives a surprising amount of comfort, simply knowing that your condition probably isn't related to one of these issues but is in fact related to something that is potentially reversible.

Yes I am seriously considering going into scientific research to study this. As I have a science background, but I'm not sure how easy it would be to get funding for research or if anyone would even allow you to. I can honestly envision, treating this in a sort of underground lab experiment (which could be very feasible).

I'm fairly convinced that this relates to changes in gene expression. If you have the time, read my post in the Theories of PAS section on the 5 alpha reductase theory. You could apply the same logic I used in this theory to any other genes but the 5AR1 gene seems to be the most likely candidate. The bottom line is Accutane does and can change gene expression. This is a fact, its already been proven in studies of the skin. We also know that Accutane works in the brain as well, given the large number of retinoid receptor in the brain. Now here is where things get interesting. All the cells in your body change over time and renew, I believe every cell in the body changes every 7 years or something. So you are biologically a different person every 7 years. The only cells in the body where this isn't true, is the neurons in the brain. You have the same neurons for life. They don't renew and you may not grow new ones. Given this, if Accutane changed gene expression in certain neurons in the brain, these genetic changes are likely to persist after finishing treatment. Which seems to be the case, given the persisting side effects.

The bottom line is, you absolutely can enhance the expression of a gene using a system such as CRISPR/CAS9. Its called gene activation. We certainly will be using gene therapies for diseases of the brain, many companies are already developing gene delivery systems for the brain as there is a great incentive to treat neurological diseases with gene therapies.

If our problems are caused by changes to gene expression, if you could target and specifically reverse those changes in gene expression. There really is the possibility of reversing this condition. In fact, you could actually go far further and greatly enhance libido etc. well above normal levels using this kind of treatment (at least I think you could).

You've had PAS for 31 years? Thats a long time, I'm around 11 years now. I highly advise you take some action to try and improve your symptoms. I can recommend things which have genuinely helped me, though they may be controversial. Ultimately, the one reassuring thing in all of this. Is no matter how much of a nightmare it may feel like, you could always be dead. There is always that alternative. Given that, and given that this has happened and you can't turn back time, it makes no sense to be pessimistic or let this condition impact your life anymore than it probably already has.
 
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flynn

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Feb 28, 2018
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#8
Why is it that the PFS group is so well organised, and the syndrome recognised, when PAS isn’t?
Its a very good question and is the reason I made this forum. It seems insane there isn't a central forum dedicated to this problem. I think part of the problem, is that most people suffering from PAS are younger than those with PFS and so are less likely to organise as well as less likely to have money/resources to organise research. Additionally, I believe the head of the PFS foundation who is a doctor, lost his son to PFS. So given his job and history he was able and had a strong incentive to set up an official foundation.

The funny thing is, I bet there are far more people out there suffering from PAS than PFS. It's just that either people haven't connected their symptoms with Accutane use or they are too apathetic to do anything or they are scattered around different forums. In honesty, I don't say this to simply promote the forum for my own gain. But I really believe it would massively benefit the community and any efforts to organise and treat this condition by getting people over to this forum or an improved version which is dedicated to PAS. If you have are in contact with anyone with PAS. It would be great if you could send them a link to the forum.
 

tom.math

New member
Mar 10, 2018
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#9
Crazy or not...seems like the most positive/likely thing I've come across on my trawls of the web. Impressed, Flynn, by your optimism. I'm not a scientist: are we saying then, that the epigenetic changes from Accutane are reversible in theory?

Several years for a therapy though, even if they can find one...it's been 31 for me, not sure how many more I can take!
thats a long time man! did you report your side effcts? I'm only a few years in. feel optimistic though
 
Jun 26, 2018
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#10
I don't really know much about this stuff but gene changes sound very worrying to me anyhow, i did some googling around and found that they can't target certain genes in a specific spot, it has to be a body wide thing which would more than likely cause some sort of cancer or other mutations.
Is it possible that up or downregulated genes can reverse themselves without using a gene edditing tool, could a researched medication fix the altered gene expressions? I'm just curious because how would some people recover and some don't.
Sorry for seeming negative, im just really scared about the idea and hope it's something not as deep as genes.
 

Dubbya_B

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Jun 27, 2018
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#11
I don't really know much about this stuff but gene changes sound very worrying to me anyhow, i did some googling around and found that they can't target certain genes in a specific spot, it has to be a body wide thing which would more than likely cause some sort of cancer or other mutations.
Is it possible that up or downregulated genes can reverse themselves without using a gene edditing tool, could a researched medication fix the altered gene expressions? I'm just curious because how would some people recover and some don't.
Sorry for seeming negative, im just really scared about the idea and hope it's something not as deep as genes.
Hi Devolution,

The great thing about CRISPR/CAS9 systems is that they excel in targeting a certain gene in a specific spot. The CAS9 enzymes used when this technology was emerging often cut near sites with sequences similar to the target. For instance, they would sometimes cut adjacent to a sequence where only 19 out of 21nucleotides were complementary to the guide RNA. You would end up with the introduced gene being inserted into multiple sites in the genome. Newer systems have been implemented that utilize CAS9 enzymes with exact specificity.

Cells with dysfunctional p53 tumor supressor (involved in apoptic signalling that causes a cell to commit suicide if it is badly damaged or out of control with growth) are far more likely to be transformed by CRISPR. That is, cells with functional p53 are more likely to self-destruct due to the process of integrating a CRISPR- introduced gene and cells with dysfunctional p53 are more likely to continue growing and replicating after a successful integration.

Cells with dysfunctional p53 were inadvertently being selected in the early embryonic stages because they were positive for the CRISPR-introduced gene. This led to organisms in CRISPR studies being selected that lacked p53 and thus, had a higher rate of tumor development and cancer, but it was not the CRISPR gene-editing that caused this. Some bloggers and news sites picked-up on studies describing this association and erroneously interpreted it as meaning "CRISPR causes cancer".

There are all sorts of drugs and substances that have been found to alter expression of specific genes. Accutane itself has been shown to do this during treatment and CBD has been found to persistently down-regulate the keratin gene.

We need to learn more about what happened to us before we can treat it. That's the bottom line.
 
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Dubbya_B

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Jun 27, 2018
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#12
Wanted to share these studies describing CRISPR systems that use deactivated CAS9 to induce epigenetic changes. No one can say that it can't be done, or that it hasn't been done yet, any longer!

Thanks to axolotl over at propeciahelp for finding these:

H.-K. Liao et al., “In Vivo Target Gene Activation via CRISPR/Cas9-Mediated Trans-epigenetic Modulation,” Cell, vol. 171, no. 7, p. 1495–1507.e15, Dec. 2017.

X. S. Liu et al., “Editing DNA Methylation in the Mammalian Genome,” Cell, vol. 167, no. 1, p. 233–247.e17, Sep. 2016.
 

BD_Acc

New member
Mar 11, 2018
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#13
Thanks for the response. It's the main reason I made this forum, as so much of the talk regarding this syndrome is merely people complaining and wishing it had never happened. People also make pretty big assertions without really thinking about the causes without providing much evidence or thought about the likelihood of that theory. The reason I made the theories of PAS, is to present each theory and weigh it up. I honestly think some of the darker theories as to how PAS arises relating to schizophrenia or brain damage, don't hold as much weight as people like to think. And it gives a surprising amount of comfort, simply knowing that your condition probably isn't related to one of these issues but is in fact related to something that is potentially reversible.



You've had PAS for 31 years? Thats a long time, I'm around 11 years now. I highly advise you take some action to try and improve your symptoms. I can recommend things which have genuinely helped me, though they may be controversial. Ultimately, the one reassuring thing in all of this. Is no matter how much of a nightmare it may feel like, you could always be dead. There is always that alternative. Given that, and given that this has happened and you can't turn back time, it makes no sense to be pessimistic or let this condition impact your life anymore than it probably already has.
What's the stuff which has improved your symptoms so far? Don't really care how controversial. I've tried some weird shit.
 

Lost

New member
Mar 10, 2018
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#14
@BD_Acc bro try bupoprion. This is the only thing that has produced something for me. It restored a very little bit of sensitivity to my penis.

If you need prescription, go to a psychiatrist and tell them about your condition. They will prescribe some SSRI like fluoxetine or somehting like clonazepam at which point you say you cant take any SSRI because PSSD has the same side effects. Then they will prescribe bupoprion since it doesn't cause sexual side effects.
 
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BD_Acc

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Mar 11, 2018
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#16
@Lost thanks man. I probably won't try it because I'm not a big fan of anti-depressants as a drug class for a few major reasons, but I'm definitely going to look into it.