Playing with Viruses

This post may end up being a big deal. Or, it might not.

This month I have learned how to grow my own bacteriophage for dirt cheap. I have also been dedicating as much time as possible to playing with genetic circuits and simulations, and attending several talks on CRISPR.

Myself and my close friends will be starting our first trials of homemade viral therapy on ourselves within the next six months. I plan on documenting everything so that we can make tutorials and share them with the community.

Now, the REALLY cool part: Gene therapy is stuck in the mud a bit right now. It has been hard to reach significant rates of infection in somatic cells. If you take a pill or inject virions into your muscles for example, you are going to experience mosaicism, or a difference in genotype among your cells, as relatively few cells will be effected.

My team and I may have discovered a way to make gene therapy 100% effective across the board. This is the specific reason I hope to edit my genome within the next three months. If we can make a viral therapy 100% effective and distribute it openly, no government in the world can stop us, and we could see a world where our own microbiomes become our in-house security systems, by editing our bacteria as well as our own somatic cells to wage war like never before on harmful pathogens and cancers.

Will be posting updates as things progress. End goals are to have easy tutorials that even a ten-year-old could follow, and at least three significant edits done to myself by the end of the year, as the series progresses.. We can talk about whether arming children with biotechnology is a good idea somewhere else though, haha.

Comments

  • edited March 2018

    How did you grow bacteriophages?

  • I transformed E. Coli with plasmids coding for virion proteins and put a LacZ promoter on them so I could selectively only produce viruses when I added IPTG, which acts like a non-digestible analog to lactose, so you can use it to turn bacterial genes on fairly easily. I am finding purification of the viruses to be little tricky though.

    Definitely looking to improve my methods in the future, which ties into my idea to make stable cells that produce virions without lysing the cell; it would be far more efficient.

  • Sounds pretty great. And the world moves one step forward towards making me glow in the dark...

  • How did you achieve 100% effective infection? How did you measure this effectiveness?
    If you truly did this, then this is worthy of a great publication.

    I would love to hear what you did and what you're thinking of doing. Maybe I can help some. I am considering putting together a group where I am as I have quite a bit of equipment and help is always favoured. Anyways, maybe we could work together on making gene therapy more effective.

  • @RealityWizard As soon as I get things confirmed, I am planning on making a huge announcement. In a nutshell though, I've designed a genetic circuit that allows cells infected by a lysogenic virus to sustain production of more viruses at a very controlled rate, allowing even single cell to be the foundation of a potential full-tissue transformation. It has been working in colonies but I need to do further testing. I am kind of scrapped for cash and trying to come up with ways to fund this project right now, admittedly.

  • Adenovirus might be worth looking into
  • So your big breakthrough is that you're not using replication deficient viruses and you're hoping you can control it? Am i understanding that right?

    All that'll do, assuming it doesn't just make you sick, is make for a large immune response and the gene you tried to deliver will get chewed out by your immune system, assuming it doesn't give you lupus. The whole point of non replicating viruses is that it stops spreading after application. That gives you a lot more control over dosage which is important because viruses do have a cytotoxic dosage. Whereas if it's still replicating you can run into weird toxicity issues, or too many copies of the gene in a single cell which can make it overtired or kill it.

    If the issue is coverage then there's plenty of ways to get better coverage without letting a virus run rampant n your system. Lots of small doses being the classic option.

  • @chironex The breakthrough is doing a large number of small doses by scheduling activation of the viruses to be very selective, as well as limit the production. I have been able to control the cycles very well so far in bacterial colonies alone. By doing this scheduling, I can avoid cytotoxic dosage completely.

    The problem with non-replicating viruses is the need for lots of small doses, which even then fail to accomplish the job, let alone being particularly unappealing to anyone not diving headfirst into biohacking.

  • Sooo excited. Can you email me, [email protected].

    I want to utilize Crispr on myself, but I’m not sure how to carry out the experiment. I read a lot, and know what’s its asking, but I have little confidence in performing it myself.

    Also, I have some ideas. You can control the expression of Cas9 with UV ligth according to a study, and you can lower or turn of cell mediated immune responses to allow Crispr to work before your body develops anti-bodies against it, if it doesn’t already have them.

    I read about Crispr weekly, if not daily.

  • Oh for fucks sake. If you don't know what you're doing, you shouldn't even consider using crispr on yourself. Hell even if you do, you probably still shouldn't. The professionals that DO know what they're doing are the first to tell you not to do that. Crispr is brand new tech and every week more papers come out showing we don't know enough about how it works and risk of unintended mutations can be very high without proper thought, and even if things have been carefully considered because of the complexity of biology. It's cute that you read about crispr, but evidently you haven't taken a fucking biology class. Jesus.

    I genetically modified myself because I knew what I was doing. Note that I did so with access to a 50k+ mammalian cell culture lab and many trained professionals to help me as well as having years of biology school and lab experience behind me. For fucks sake, genetic modification, especially of yourself, is not something to be taken lightly. Specially if you're doing it for fun. GAHHHHHH

  • @chironex said:
    Oh for fucks sake. If you don't know what you're doing, you shouldn't even consider using crispr on yourself. Hell even if you do, you probably still shouldn't. The professionals that DO know what they're doing are the first to tell you not to do that. Crispr is brand new tech and every week more papers come out showing we don't know enough about how it works and risk of unintended mutations can be very high without proper thought, and even if things have been carefully considered because of the complexity of biology. It's cute that you read about crispr, but evidently you haven't taken a fucking biology class. Jesus.

    I genetically modified myself because I knew what I was doing. Note that I did so with access to a 50k+ mammalian cell culture lab and many trained professionals to help me as well as having years of biology school and lab experience behind me. For fucks sake, genetic modification, especially of yourself, is not something to be taken lightly. Specially if you're doing it for fun. GAHHHHHH

    Accidental making myself shoot lasers out of my eyes!

    Because that’s totally how that goes in comic books. :P

  • I understand that's meant as a joke but there have been a lot of people recently expressing a kind of, " I'm ok if I fuck it up and die" mentality. I really don't like animal testing. I'd love it if we could totally avoid it in this community.. but realistically.. wouldn't you like to see the effects in say.. three generations of rats before considering humans?
  • @Cassox said:
    I understand that's meant as a joke but there have been a lot of people recently expressing a kind of, " I'm ok if I fuck it up and die" mentality. I really don't like animal testing. I'd love it if we could totally avoid it in this community.. but realistically.. wouldn't you like to see the effects in say.. three generations of rats before considering humans?

    I'd say yes, and at the same time a big no... three generations of rats later we would realize they're not human. Do you know how many stem cells trials have worked perfectly on Mouse, pigs, yet filled when given to humans.

  • Yeah. I agree. I don't know of a good alternative method
  • I've been experiencing some difficulties getting GFP genes to spread through mammalian cell cultures using this method so far, but I am going to try a variety of vectors once I get access to a sequencer. I want to see exactly what each viral vector is doing in detail, and since I can't easily tell why my experiments are failing, I don't want to dump more resources into it just yet.

    @Cassox I'd say there really is no alternative to testing on humans, but simulations and cell cultures can have been helpful. I can't recommend testing on human fetuses of course, but scientifically it is the most useful direction to go. CRISPR researchers have already gotten hell even though they terminated the fetuses at the legal time. Maybe in China?
  • edited August 2018
    It's funny that you should mention China. This is an older article but it does state that we have already lost some of our greater more prominent stem cell researchers to Asia. Any of you that is interested in that article here's the link
    https://www.humboldt-foundation.de/web/1255.html
  • > @IvoTheSquire said:
    > @chironex said:
    > Oh for fucks sake. If you don't know what you're doing, you shouldn't even consider using crispr on yourself. Hell even if you do, you probably still shouldn't. The professionals that DO know what they're doing are the first to tell you not to do that. Crispr is brand new tech and every week more papers come out showing we don't know enough about how it works and risk of unintended mutations can be very high without proper thought, and even if things have been carefully considered because of the complexity of biology. It's cute that you read about crispr, but evidently you haven't taken a fucking biology class. Jesus.
    >
    > I genetically modified myself because I knew what I was doing. Note that I did so with access to a 50k+ mammalian cell culture lab and many trained professionals to help me as well as having years of biology school and lab experience behind me. For fucks sake, genetic modification, especially of yourself, is not something to be taken lightly. Specially if you're doing it for fun. GAHHHHHH
    >
    >
    >
    >
    >
    > Accidental making myself shoot lasers out of my eyes!
    >
    > Because that’s totally how that goes in comic books. :P

    ..you can have the eyes..I want wolverine's healing... Lol
  • CRISPR is a great tool but it is hard to control in some organisms. Human testing, from China specifically, is showing it is not as site-specific as it needs to be. Even in yeast it can cause mutations in other areas of the genome or even insert in the wrong place. If you have an alternative or modification to the system that has given it better site-specificity I would be super interested...well most of the molecular community would be...but me too! This is the same problem I'm having with my project. I"m not sure how to make it more specific for humans and it might not be the best answer for us. We might need to develop an alternative.

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