[DIYbio] Re: are we at the dawn of a self experimentation gene therapy trend?

I do agree that PEPCK-C muscle overexpression is a promising target for "long lived super strong transhumans". Beyond that, I have to argue the rest of your points.

The 'mighty mice' study you reference used germline genetic modification, followed by inbreeding, to generate the actual supermice. As they mention, the initial pre-inbreeding mice, with levels of the enzyme already much higher than wild-type, did not act differently from their wild-type counterparts. Now, a germline mod implies that every muscle cell in these founder lines had perhaps one copy of the transgene, albeit with the weak-ish (versus CMV) skeletal actin promoter. Nonetheless, that's already a hell of a lot of transgene being expressed, with no noticeable effect until they were bred to produce descendants with even more copies of the transgene. You can't dream of getting anywhere near that level of expression with postnatal delivery, especially non-viral.

Muscle electroporation is relatively efficient compared to e.g. chemical methods, but as with all plasmid gene therapy, works far better in mice than it does in humans. Even if you can develop a delivery system that is tolerable to the subject and doesn't kill a significant fraction of their muscle over the thousands of injections necessary, your expression levels won't be high enough to make a significant dent in the levels of PEPCK. From the activity units-to-milligram conversions I've seen for PEPCK, the 'mighty' mice were hitting several milligrams PEPCK protein per gram of muscle, and possibly far more; I'll have get a more concrete number.

People with far more resources than us have been trying to get plasmid electroporation in muscle to work for over 20 years, and so far no one's had much success. There's a few edge applications where it's efficacious enough - DNA vaccines don't produce much antigenic protein but it's enough for an immune response if you jam enough adjuvants in there, or a protein like EPO, which has such a low effective dose that you might get therapeutic benefit. However, reaching even milligrams of protein per kilogram of body weight isn't there with plasmids, much less mg/g, and not for an extended duration, so you're gonna be constantly getting electrocuted just to keep the levels up.

Okay, on to the plasmid you linked. Certainly one "major challenge has been a suitable vector". This is not "potentially exactly that vector". It's just a generic mammalian expression vector that someone wrote a bunch of question marks on. It'll probably work okay on cells in a dish but in vivo is an entirely different matter. The bacterial part of the cassette (resistance gene and origin) is strongly silencing in mammalian cells, and the origin has additional problems since it has strong secondary structure, making the plasmid unstable during and after delivery - see e.g. the development of minicircle vectors that cut out all the bacterial elements, and/or the addition of insulator elements like S/MARs to block the silencing.

There's plenty of other stuff wrong with the plasmid if you wanted to even try and optimize it, but if the aim is to get publicity by injecting it into yourself with no actual biological effect, then go hog wild; that seems to be the state of things right now. I can appreciate that people want to start a conversation about these things or w/e, but...

On Saturday, November 11, 2017 at 12:03:16 PM UTC-8, CodeWarrior wrote:

So some of you may have noticed the creation of the open human plasmid now being offered by the odin. If you're not aware of it you might want to go and look it up (http://www.the-odin.com/open-human-plasmid/). It is basically a plasmid for gene expression in human cells. Advertised for in vitro use but in theory just as usable in vivo. Of course getting a plasmid into human cells in vivo is quite difficult. Unless you are targeting muscle cells where electroporation has been demonstrated as an effective and efficient method for transfection of muscle. (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2975437/ and https://www.technologyreview.com/s/603217/one-mans-quest-to-hack-his-own-genes/)

Now this brings to mind an experiment I read about many years ago. (Biochimie. 2008 June ; 90(6): 838–842. and J Biol Chem. 2007 November 9; 282(45): 32844–32855.) In short a genetic modification of mice causing them to express a protein (PEPCK-C) normally expressed only in the liver and fat in muscle instead had 3 major effects:

1. increased life / health span and improved peri-mortem health.
2. Increased muscle mass, strength and endurance.
3. Hyper sexuality and aggression (sexuality and aggression are known to be closely linked in mice).

It's often been speculated by amateurs and transhumanists that replicating this work in humans might lead to long lived super strong transhumans. The major challenge has been a suitable vector for human somatic muscle. I know many will wish it wasn't pointed out but the open human plasmid in combination with electroporation is potentially exactly that vector. The electroporation targets muscle quite specifically as I understand it where as the open human plasmid will easily accept the PEPCK-C coding region which will be strongly expressed in muscle by the CMV promotor.

Now here I must give a disclaimer. I'm not a lawyer, this is not legal advice. However I believe self administration of this process is legal in the UK. In the UK genetic engineering is mostly regulated by the GMO regulations that relate to creation of genetically modified organisms. To be a genetically modified organism you have to be a non human biological organism or capable of transferring genetic material into one. The plasmid isn't capable of self transferring itself into cells. If it were a GMO every plasmid or DNA section ever manipulated in a lab would count as a GMO as they could all be transported into muscle cell through electroporation. However no human can be considered a GMO they are specifically excluded under the wording of the regulation. So it should be posable to administer this therapy to humans with out violating the GMO regulations. Of course there are other regulations relating to unlawful human experimentation but I expect most of them would be mitigated by only experimenting on ones self.

So what do you think are we on the verge of an explosion of self creating transhumans? You'd have to be pretty brave to strap your self down, inject your muscles with something you made from something you bought online, jab needles into a large portion of your body, then run very significant electrical currants through your self.

Now aside from the feasibility of this, which I'd be eager to hear your views about as well. I'd be interested to hear if anyone thinks amateurs are likely to go this far with self experimentation? And if this sort of thing catches on what then?