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DMT and genetic engeneering Options
 
shago
#1 Posted : 12/10/2008 3:23:08 AM

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Do you ever dream about a plant containg extreme amounts of DMT per weight? what about a plant containg 3%? sounds good, isn't it?

Well, today this dream can be made true with genetic engeneering as more researsh is done in the field. This is the century of the biology and genetics.

We all have heard about modified plants and crops. For example modified canabis plants.
Would you try a modified mimosa hostilis? Or will you hold on to traditional stuff?
Have you heard about such experiments going on?

Let us share some points of view.

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jamie
#2 Posted : 12/10/2008 4:07:36 AM

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There is alot of ppl working on the issue of upping the alkaloid content of DMT containing plants. especially when it comes to Phalaris species..same with cactus..lots of hybridizing going on as we speak..

I think coatl might know more..Ive seen him post on the issue of "superstrains" more than once..



 
timeloop
#3 Posted : 12/10/2008 4:58:45 AM

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quote from strassman in: "Inner paths to outer space - journeys to alien worlds through psychedelics and other spiritual technologies"

"...scientists have isolated the gene that synthesizes the DMT-forming enzyme in humans and have inserted this gene into a virus. After they infect mammalian cells with this virus the cells produce DMT in the test tube"

from"
M. A Thompson, E. Moon, U.-J, Kim, J. Xu, M. J. Sixiliano, and R.M. Weinshilboum, "Human Indolethylamine N-methyltransferase: cDNA Cloning and Expression, Gene Cloning, and Chromosomal Localization," Genomics 61 (1999): 285-97

 
jamie
#4 Posted : 12/10/2008 6:27:50 AM

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now thats interestingVery happy , i'l sign up for that vaccine!
 
Infundibulum
#5 Posted : 12/10/2008 10:33:04 AM

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As the most authoritative person on molecular genetics and genetic engineer around here I would say that it is quite complicated if it ever works. It can seriously takes a lot of years of intensive work (maybe 10 but at least 5). We have plenty of gaps in the dmt synthesis pathways and how they are utilised from the plants so that we cannot build a coherent solid strategy to meet such goals. No talk about the funding either.

If one wants more, then extract more material, it is so much simpler. Mimosa can already have at around 2% active alkaloids, phalaris brachystachys on a recent claim can have up to 3% actives.

How much more do you want you greedy people, 10-20%? The plant itself may not even be able to cope/survive with such amounts!

The strassman quote is also a bit misleading by making people think that some super scientific breakthrough has been achieved. In my world this actually means nothing more than "one of the genes in the dmt synthesis pathway has been isolated". Nothing really more. It is more or less a standard technique that me and my colleagues use on daily basis and we do (and can do!) with every other gene from every other organism. The inclusion of the word "virus" from strassman seemed to make things more complicated by making easier for people to entertain the idea that "one can get an injection of the virus and get himself producing dmt somehow"

But this is far from truth...

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deedle-doo
#6 Posted : 12/10/2008 2:51:21 PM

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shago wrote:
Do you ever dream about a plant containg extreme amounts of DMT per weight? what about a plant containg 3%? sounds good, isn't it?

Well, today this dream can be made true with genetic engeneering as more researsh is done in the field. This is the century of the biology and genetics.

We all have heard about modified plants and crops. For example modified canabis plants.
Would you try a modified mimosa hostilis? Or will you hold on to traditional stuff?
Have you heard about such experiments going on?

Let us share some points of view.


This come up from time to time. For plants breeding and selection are your best bet. If you really wanted to engineer a stable population of DMT producing plants it wold have to be arabidopsis thaliana. There are good kits for arabidopsis transgenesis that anyone can use who can read and follow directions.
If you were fine with transient DMT production you could infect a plant of your choosing with a virus that will give great bulging tumors that express your transgenes.

BUT, for this kind of thing you should really be using brewing yeast. There are idiot-proof kits for multiple transgenesis in saccromyces that pretty much always work. Depending on how many transgenes you needed to add (I'm guessing 4-5 but who knows, won't know for sure untill it is tried) this would only cost 2000-4000USD and could be done in a couple of months. Once you had the yeast strain you could lyse the cells, pellet down and do an extraction on the culture medium to obtain the product.

Fun to think about but not very practical. Better just to get yourself set up for large scale plant extractions like Infundibulum advised.

 
Infundibulum
#7 Posted : 12/10/2008 3:14:42 PM

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deedle-doo wrote:
BUT, for this kind of thing you should really be using brewing yeast. There are idiot-proof kits for multiple transgenesis in saccromyces that pretty much always work. Depending on how many transgenes you needed to add (I'm guessing 4-5 but who knows, won't know for sure untill it is tried) this would only cost 2000-4000USD and could be done in a couple of months. Once you had the yeast strain you could lyse the cells, pellet down and do an extraction on the culture medium to obtain the product.


One could not even have to lyse the cells. With a wee bit of extra work the dmt pathway enzymes could be targeted in the secretory vacuoles of the yeast. This would be far more practical. So you brew everything like you would brew beer (which is dead simple BTW), and the yeast instead of fermenting any beer, "spits" out dmt in the brew.

One can then go straight and drink the brew aya-style or simply basify it and extract it.

The cop asks "what's inside there", you reply "Oh, it's my home brewed beer in progress". The cop asks "what are those chemicals" you reply oh, "Oh, it's the stuff for making the fermantation brew, to add taste or even sterilise the barrel". And you wouldn't be more honest about your answers.

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deedle-doo
#8 Posted : 12/10/2008 3:34:46 PM

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Infundibulum wrote:


One could not even have to lyse the cells. With a wee bit of extra work the dmt pathway enzymes could be targeted in the secretory vacuoles of the yeast. This would be far more practical. So you brew everything like you would brew beer (which is dead simple BTW), and the yeast instead of fermenting any beer, "spits" out dmt in the brew.

One can then go straight and drink the brew aya-style or simply basify it and extract it.



eeewwww!!! I'm not drinking saccromyces transgene maintanance culture medium. Allthough if we used a transgenesis strategy that avoided antibiotics I'm sure it would be quite healthy for the body.

Maybe if we're really lucky the transgenes will be stable even without selection and we could actually use these strains to brew beer. That'd be awesome! I'd brew an IPA. TONS of hops. Add in a b-carboline. Call it 'extra-special spiced ale.' Great for camping, not so much for tailgate parties Smile
 
Infundibulum
#9 Posted : 12/10/2008 4:36:50 PM

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I would use a low fermenting yeast anyway so it would sediment. In this respect, the brew would have an ale-style. IPA is certainly a kick ass brew. But I would save my hops and malt for a real beer brewing.

I have been brewing beer just using sugar in the past, no malt ho hops at all. It is fairly easy, but the end product does not taste amazing. It does the job however, you can get pretty drunk. So for the purposes of growing this specific transgenic yeast, no culture media are really necessary. But mind you, I find the taste of YPD broth very agreeable.

This friend of a friend actually finds the generation of this transgenic yeast a very attractive idea. Too bad he doesn't have enough money in his grant to justify buying good yeast for transgenesis, vector(s) and possibly antibodies. The rest are easy to do, primers are cheap as fuck, restriction enzymes abundant and PCR reagents always fresh so cloning the genes should be piece of cake.

Maybe he can send the plasmids (should he actually does the initial cloning in an easy bacterial vector like P-GEM T-easy from promega) to some other person to continue the job?

???

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burnt
#10 Posted : 12/10/2008 5:27:19 PM

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genetically engineering organisms to produce more of a specific alkaloid is tough as heck. there are many promising cancer drugs currently facing this problem. none the less its an interesting approach. of course DMT is much simpler and many organisms out there would only need one or two genes to do it. however you also have to think about feedback regulation genes etc etc....

however breeding is probably easier and just as effective (especially without a few grand in research grants!).

also dmt is relatively cheap and easy to synthesize on an industrial scale (compared with insanely comlicated structures found in some cancer drugs) that its not worth funding such a project.

there is also one other issue. DMT is toxic to some organisms (some kinds of livestock) and possibly other critters, it may also be toxic to the plant itself, so one may not strains cross breeding with wild grasses etc. sometimes when people try to get plants to make lots of alkaloids the alkaloids kill the plant or cell culture. althought I dont think there is any evidence that dmt is toxic to plant cells (remember alkaloids are often stored in various cell compartments etc to keep plant cells safe, unless they are under pathogen attack) it could be.
 
Infundibulum
#11 Posted : 12/10/2008 6:37:19 PM

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I hear you, you're absolutely right. As far as the yeast system is concerned, excretion of the end product (dmt in this case) and compartmentalisation of the introduced proteins has been shown to be promising in overcoming said problems. The latter is important and has been a saviour (especially to protein crystalographers, who, in need for massive amounts of the x, y, z protein they overexpress in yeast find out that the protein itself is highly toxic).

I am not concerned too much about the negative feedback loops that are so abundant and can limit gene expression. The transgenes are going to be under the control of some strong constituvely active (or even inducible) promoter so that large amounts are made overcoming host regulatory pathways.

I have worked in the past with similar promoters, more specifically the AOX-1 gene promoter that is massively induced by the presence of methanol in the growing medium. So cloned genes could be insanely overexpressed if some methanol was added in the yeast's medium. But that was in the yeast Pichia pastoris. I am fairly confident that things have moved since then and very good vectors can be bought (or constructed) to do this type of work in Saccharomyces cerevisiae.

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timeloop
#12 Posted : 12/10/2008 11:41:40 PM

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Infundibulum wrote:

The strassman quote is also a bit misleading by making people think that some super scientific breakthrough has been achieved. In my world this actually means nothing more than "one of the genes in the dmt synthesis pathway has been isolated". Nothing really more. It is more or less a standard technique that me and my colleagues use on daily basis and we do (and can do!) with every other gene from every other organism. The inclusion of the word "virus" from strassman seemed to make things more complicated by making easier for people to entertain the idea that "one can get an injection of the virus and get himself producing dmt somehow"

But this is far from truth...


considering that strassman mentions:

Quote:

...After they infect mammalian cells with this virus the cells produce DMT in the test tube...


wouldnt this indicate that there has been a little more effort put into the research than simply isolating one of the genes in the dmt synthesis pathways. Seems to me like this is something they have actually succeeded in successfully experimenting.

Although it is highly unlikely that we can infect ourselves with a virus and have DMT produced in every cell in our body...

SWIM has had some elves wisper to him that the evolution of humankind hinted at around 2012 is actually precipitated by this virus infecting everyone on the planet and transporting us all into hyperspace permanently!

Laughing

can someone please confirm the likelyhood of any phalaris grass containing 3% nn-DMT? or would this be a mixture of other alkaloids... SWIMs brief research into phalaris was ended after it was discoverd that there are an amount of other unwanted alks that would have to dealt with.
 
fourthripley
#13 Posted : 12/10/2008 11:56:05 PM
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Quote:
can someone please confirm the likelyhood of any phalaris grass containing 3% nn-DMT?


The 3% figure is in relation to P. Brachystasis and is now pretty much accepted as a typo, misplaced decimal point. I quite believe 0.3 might be achievable.
mistakes were made
 
Infundibulum
#14 Posted : 12/11/2008 1:50:36 AM

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timeloop wrote:
wouldnt this indicate that there has been a little more effort put into the research than simply isolating one of the genes in the dmt synthesis pathways. Seems to me like this is something they have actually succeeded in successfully experimenting.


Not really; strassman is a bit full of bullshit here. What they did was that (according to the paper in which the experiments are described in detailed:

1) clone the gene into a virus. (the "virus" is just a vector for the gene, i.e it can be used to transfer the gene into the cells which it can transfect).

2) the cells were COS-1 cells (insect cells) that after the viral transfection expressed the gene in high amounts.

3) tryptamine was supplemented into the medium in which the cells were growing and the tryptamine was methylated to dimethyltryptamine.

The "little" detail of tryptamine addition to the culture medium (i.e. the substrate of the enzyme they were overexpressing actually made them produce dmt. Should tryptamine not have been added, no dmt would be produced.

I really do not see the point of strassman publishing information in such a misleading way. His phrasing certainly gives the wrong message especially to non-specialists forcing the idea that some great scientific breakthrough has been achieved.

Strassman's credibility is sometimes at stake, at least from my point of view.

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deedle-doo
#15 Posted : 12/11/2008 7:12:38 AM

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Infundibulum wrote:

I am fairly confident that things have moved since then and very good vectors can be bought (or constructed) to do this type of work in Saccharomyces cerevisiae.


There are kits for everything now. These kits are designed to allow novices to make proteins. Anybody could do this with a lot of work, a moderate budget and a well equiped laboratory. DMT would be easy, just add one gene and supplement the media with tryptamine, in a nation or territory where this was totally legal of course.

Creating novel-target compunds via enzyme expression looks like a colossal pain in the ass. Seems like they're making good progress though. I saw a talk by a dude who studies enzymatic reactions in sea-sponges and their bacterial symbionts and uses them to produce novel compunds. this guys hangups were needing better control enzyme stoichiometry and timing.
 
Infundibulum
#16 Posted : 12/11/2008 12:46:41 PM

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The yeast can be made to produce tryptamine in the first place, maybe this could be the first goal to achieve. And tryptamine does not seem to be toxic for yeast. The gene that could be inserted is NM_016672 (from pubmed; it's from mouse, shouldn't matter too much though) which can decarboxylate tryptophan and phenylalanine. And it's almost 1500 bases long, so cloning, subcloning and transfection are going to be pretty much routinely.

The next one is the indoleamine N-methyltransferase, again form mouse, check pubmed for NM_009349. It is just less than 800 bases long, so it won't too much of a hassle to work with.

But of course, all these things are totally hypothetical, no-one should try such things without obtaining a licence for performing the said activities. I am serious and I would be very very afraid to do such things, this goes without saying!

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lorax
#17 Posted : 12/11/2008 2:15:57 PM

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why not just synthesize it? i guess that'll be much easier than going through all the trouble of potentiating specific plants.
I am the Lorax. I speak for the trees. I speak for the trees, for the trees have no tongues. And I'm asking you, sir, at the top if my lungs.. (all posts are fictional and are intended for entertainment purpose only)
 
Infundibulum
#18 Posted : 12/11/2008 2:29:24 PM

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Because organisms do it so much better than chemists. For synthesis one needs a decent lab to perform the procedures involved. And you'll have to go through the process every time you require some. With genetic engineering you need a lab and some good time until you create the transgenic yeast/bacteria/plant etc, but as long as you make it that's it! It can be distributed to everyone and everyone will just have to grow a simple organism to get spice.

The enzymes can perform chemicals reactions with an accuracy, specificity and efficiency that many chemists would envy. Having an organism to do it for you is the best one can come across for acquiring spice. Not everyone can do synthesis but everyone can brew some yeast or grow a plant.



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shago
#19 Posted : 12/12/2008 1:01:26 AM

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more funds should be given to investigation in entheogens plants and hallucinogens, the prohibition stopped the progress in research. I think this substances have many medical usages like in psychology. And many are used as sacraments. I think that people that use them for religious purpose won't be using trans genetic or modified plants, because many persons believe this is violence against nature.

Can someone tell me if modified food could result harmful for the organism like developing cancer in the future?

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deedle-doo
#20 Posted : 12/12/2008 1:56:13 AM

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shago wrote:

Can someone tell me if modified food could result harmful for the organism like developing cancer in the future?


No cancer risk at all unless you foolishly modify the crop to make a carcinogen. It is more possible that a person could develop allergies to the modfified crop but this is probalby pretty unlikely.

Modified food cropps are not to be feared because they're unhealthy. On the contrary, many are specifically engineered to be more nutritious. You should fear modified food crops beacuase they are clonal. A whole crops worth of seed with very little or no individual variation. Then this massive colony may spread their pollen far and wide and remove the diversity from others of its kind.

We can try to engineer these plants to die after some time and not reproduce but it wont hold forever. That's why I think these plants should only be grown underground or in very remote and starving areas.
 
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