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DIY Genetic Engineering & DMT production Options
 
Attention All Shipping
#1 Posted : 6/19/2009 1:31:24 PM
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" I isolated chickpea DNA using non-iodized salt, shampoo, meat tenderizer, and a salad-spinner for a centrifuge,":http://hplusmagazine.com/articles/bio/darning-genes-biology-homebody

I've speculated in the past about taking genes from mimisa hostilis (etc) and putting them into something else, less tropical, more easily disguised, quicker growing; or of modifying it to produce DMT in leaves or in greater quantities; but I've always assumed it'd be something that's impossible to do without millions of pounds of fancy gene equipment & lots of education. From the above link it looks like DIY genetic engineering is possible & that, if not now, but shortly the possibility of modifying DMT producing organisms won't be out of reach to non-professional enthusiasts, albiet a high level of education will probably still be necessary.
 

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Infundibulum
#2 Posted : 6/19/2009 2:50:23 PM

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Attention All Shipping wrote:
" I isolated chickpea DNA using non-iodized salt, shampoo, meat tenderizer, and a salad-spinner for a centrifuge,":http://hplusmagazine.com/articles/bio/darning-genes-biology-homebody

I've speculated in the past about taking genes from mimisa hostilis (etc) and putting them into something else, less tropical, more easily disguised, quicker growing; or of modifying it to produce DMT in leaves or in greater quantities; but I've always assumed it'd be something that's impossible to do without millions of pounds of fancy gene equipment & lots of education. From the above link it looks like DIY genetic engineering is possible & that, if not now, but shortly the possibility of modifying DMT producing organisms won't be out of reach to non-professional enthusiasts, albiet a high level of education will probably still be necessary.

Nah, it's not that easy to do genetic engineering. Isolation of DNA btw is far easier than using "non-iodized salt, shampoo, meat tenderizer, and a salad-spinner for a centrifuge". Actually one needs just water and sodium hydroxide and a heat source. The big challenge is to isolate the genes of interest and clone them into an organism.

SWIM as well as other SWIMs around have the knowledge, the funds and the materials but not the time to isolate genes and make genetically modified organisms that produce dmt. But it is a seriously lengthy project to do that may take from 1 year (if one's really lucky) to 10 years. It's not something that one can do clandestinely in the afternoon after work.

But FYI, there had been some past discussions for this issue on the Nexus. A nice way seems to be the generation of transgenic brewer's yeast to express the right genes. Then you brew your beer and voila! it has dmt inside. One can either drink the beer with a MAOI or directly extract it from there.


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bufoman
#3 Posted : 6/19/2009 3:31:18 PM

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This would be an amazing feat but like infund said it is very expensive and time consuming and much harder than just isolating DNA. There are thousands of genes within that DNA (dep on organism). These thousands of genes are than only 1% of the sequence. I am not sure a salad spinner will help isolate the desired gene (Just kidding). One needs a lot of molecular toys to do such work. Restriction enzymes, PCR, plasmids, bacteria... But yes the point is it def could be done and it would not be particularly hard if the times and funds were available.
 
TheNtt
#4 Posted : 6/19/2009 5:12:37 PM

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I thought I read in "Inner Paths to Outer Space" that Rick Strassman and his team were actually doing this.
I'll try to find what I read...
 
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#5 Posted : 6/19/2009 5:14:05 PM

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SWIMfriend
#6 Posted : 6/19/2009 6:02:44 PM

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slidewinder wrote:
"The day someone produces a cannabis plant that secretes spice will be the greatest day this planet has ever known." - unattested



You wouldn't do it with a plant, you'd use bacteria (or yeast, as mentioned above). There are only a few genes required for converting trytophan to DMT. It wouldn't necessarily be insanely difficult, and it wouldn't take "millions of dollars." But it would take a non-trivial amount of money and quite a bit of time and work...

And even when you were all done and had a bacterium that would make DMT for you, you'd still have to culture and collect DMT--which might very well be MORE EXPENSIVE and MORE INVOLVED that simply extracting from bark...at least for the individual user.
 
Observant
#7 Posted : 6/19/2009 6:24:44 PM

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what about growing the plant cells in a bioreactor ?
I read that codeine could be produced like this.

That wouldnt even require bacteria and genetic engineering - if it worked.
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SWIMfriend
#8 Posted : 6/19/2009 6:33:11 PM

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Observant wrote:
what about growing the plant cells in a bioreactor ?
I read that codeine could be produced like this.

That wouldnt even require bacteria and genetic engineering - if it worked.


You would have to ask yourself whether such a project was easier/better/cheaper than straight out organic synthesis (the goal of such a project WOULD be synthesis). I don't think actual synthesis of DMT is that big a project...so there's no need for ultra-high-tech methods when more standard techniques would work. And still, for individual use, extractions are easy and cheap.
 
bufoman
#9 Posted : 6/19/2009 6:39:31 PM

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You could absolutely transfect a plant to produce DMT. Although this would be slightly more difficult and take longer but it would not be much harder than transfecting yeast or bacteria. It would be possible to transfect cannabis. That would be an interesting combo although it would take an additional few genes to up-regulate the expression of the enzymes to a level where it would be active via smoking. Most likely it would still have to be extracted as even 3% would not be strong enough to smoke and thus cannabis would not be the best choice. However fast growing plants which are tolerant to various climates would be useful. 2 genes would be required, tryptophan-decarboxylase and indol-N-methyl-transferase. Various co-factors such as SAMe and other methyl donors would also be required. As well as upstream enhancers and transcription factors. But an effective vector could be engineered.

This project would be very expensive maybe not millions but it would be expensive. A lot of preparation has to go into locating the genes and necessary restriction enzymes require. Also a lot of expensive equipment (PCR, incubators, and other bio-tech tools would be required)
 
bufoman
#10 Posted : 6/19/2009 6:44:06 PM

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SWIMfriend wrote:
Observant wrote:
what about growing the plant cells in a bioreactor ?
I read that codeine could be produced like this.

That wouldnt even require bacteria and genetic engineering - if it worked.


You would have to ask yourself whether such a project was easier/better/cheaper than straight out organic synthesis (the goal of such a project WOULD be synthesis). I don't think actual synthesis of DMT is that big a project...so there's no need for ultra-high-tech methods when more standard techniques would work. And still, for individual use, extractions are easy and cheap.



Ergot is in some places produced in this way. A broth containing the fungus is grown and then it is extracted to recover the alkaloids. Don't know about codeine sounds interesting though.

Of course synthesis would be easier but that is not the point we are just speculating on the possibilities. It would still be possible to do the transfection. Obviously no ones SWIMs are going to do this anytime soon. But one day this may occur, for various reasons. Environmental tolerant plants, increase alkaloid content, decrease fat levels.... various factors could be weighted to produce an optimal source plant for extractions of a variety of psychoactive compounds (in the future of course).

One day chemistry may be done by using protein enzymes as catalysts. Thus you would just put tryptophan into a reactor, add your enzymes and co-factors incubate at the optimal conditions (pH, temp, ionic strength, time) and then extract. This would reduce side products and optimize synthesis of a variety of natural compounds.
 
blizznshot
#11 Posted : 6/19/2009 9:14:12 PM
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Just for a point of reference, a group at Caltech has successfully engineered Saccharomyces yeast to produce reticuline (morphine precursor). Even though the biosynthetic route has been worked out in Papaver, the research ultimately took years and was published in Nature (top-notch scientific journal).

The reason it took so long is the fine-tuning that is involved after you have your candidate genes in a cloned form. Too much production is toxic...use a weaker promoter. Now there is a bottleneck which leads to the buildup of toxic metabolic intermediates. Etc, etc.

Not to mention cross-species codon optimization, post-translational regulatory engineering, and the possible need to introduce stabilizing elements in your gene so that the protein will remain intact in a foreign organism.

In a nutshell, if the expression of a fairly well-characterized "system" warranted a Nature publication, I'd find it tough to believe that the at-home Gene Jockey could do similar work with a relatively unknown system.

I'm not saying it's impossible, but this goes far above and beyond your typical cutting and pasting, and transfecting/transforming. That's just the tip of the iceberg...
 
SWIMfriend
#12 Posted : 6/19/2009 10:24:58 PM

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blizznshot: it all depends on the number of intermediates, each requiring a different enzyme. MANY intermediates requires many enzymes, with the problems of controlling ALL the enzymes--and also being sure your intermediates are not send down another pathway by an enzyme you DON'T want, but that's integral to the bacterium...
 
blizznshot
#13 Posted : 6/19/2009 10:33:45 PM
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Indeed! And by "system," that is exactly what I mean: the group of genes required to produce any given "metabolon." IIRC, the reticuline metabolon required something like three exogenous genes, provided the yeast were fed a precursor (I forget specifically what they used...ok, it's apparent I'll have to dig up the paper). And just to be clear, the poppy paper required the tuning of expression levels and structural properties of each individual enzyme in order to make a functional system. That's why it was published in Nature.

I wasn't trying to say it's easy, quite the contrary. I was outlining the difficulties! Trust me, I'm very aware of the technicalities of this type of work. I just tried to keep the post simple by comparing the work to literature and talking about the regulation and tuning of a system as a whole, as opposed to breaking it down enzyme by enzyme.

For DMT production from tryptophan, I believe that at least three exogenous gene would be required. And that's just hypothetical. I don't believe the absolute requirements are really known in terms of what is necessary and sufficient.
 
SWIMfriend
#14 Posted : 6/19/2009 10:42:06 PM

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Yes, it's clear that it's not simple. Not only because the one you mentioned rated a Nature article, but because a ZILLION things would start being made that way and, so far, not too many things are.

For those interested, attempts to breed higher DMT-yielding plants, or easier and better extraction methods, are likely the most practical goals along these lines.
 
 
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