The Biotechnology of Cannabis sativa

Hi everyone,

Hope you will forgive introducing myself in this part of the forum. I just wanted you all to be aware of this:

The Biotechnology of Cannabis Sativa

It's a free discourse on the ways biotechnology (genetic engineering) may be applied to increase cannabis potency and to possibly create a whole mess of novel structural phenotypes.

Most (if not all) of the genes in the THC production pathways have been sequenced and can be obtained for free on sites like NCBI's entrez.

Sam Zwenger, the author of the pdf I linked even goes on to document actual transgenic GFP (green fluorescent protein) cannabis. Doesn't look like much, but this is a milestone in DIY genetic engineering. Next year, I hope to duplicate his results and possibly start my own transgenic cannabis projects.

......good luck.....

Yeah I suppose that's the appropriate response...

Thanks though. It's going to take a lot of patience and trial and error but in the meantime I'll still be growing dank bud of the non-transgenic variety.

What I am searching for is an investor to help me set up a lab and work more discretely. In the meantime, if you have questions about modifying the plant you can send me an email, [email protected]

There are some technicalities to watch for and most likely only plant biotechnologists will be cautious/aware of. I'd like to present some data at the next Cannabis Cup, if they would have me. Thanks for the interest.

Sam

This is a killer read man, thanks! Do you have an account with this website so you could send me the .PDF?

thanks very much northernlights, been doing my own reading on the subject and others....if u have any other info on the subject and whould be willing post it whould be very kool..meangreen

hey NL do u know if the GFP colors of the modified pot plant...can be seen when looking at the genes ( if the THC is "blue" is the gene/s for THC also blue)?

MEANGREEN69 said:

hey NL do u know if the GFP colors of the modified pot plant...can be seen when looking at the genes ( if the THC is "blue" is the gene/s for THC also blue)?

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I'm not entirely sure what you're referring to here. I think you may be confusing chromosome staining (like karyotyping) with gene sequencing.



It would be hard predict the amount of expression of the green fluorescent protein in GFP cannabis as gene expression depends on many different factors.



For example, researchers have obtained very high levels of GFP expression in GFP tobacco that seem to be consistent throughout the entire plant (see below).



(note: this is most likely a long exposure shot of the plant glowing after a period of being bombarded by blue light)



However, an initial transfer of a GFP plasmid to C. sativa may not yield such stunning results until more solid expression vectors are found.


P.S. Sam Zwenger, I'm super stoked you replied to this thread! I just sent you an email

WOW!! thats bad ass...thanks for clearing is up 4 me..the only info i can get is off the net 4 now..makes more sense after reading that link...been reading alot on Isolation of protoplasts and the fusion of them as a way to cut back on breeding time....meangreen

As a person who majors in biochemistry and has done genetic engineering on bacteria in the lab, I feel that it should NEVER be applied to any thing that we consume. Not only is there not enough information on how modifying the genes of an organism that is introduced into the wild could have harmful effects on the environment. There is all ready signs of how genetic engineering of certain types of plants such as corn have actually contributed to the reduction of different varieties of corn. I rather the THC content of weed be determined by breeders NOT by scientist. You plant a seed that has been genetically engineered into the wild, some of the seeds will produce pollen that will travel around a certain region. Pollen can latch onto certain animals and travel hundreds if not thousands of miles. The pollen with the untested genes are then introduced to another line of genetics and could destroy them. Genetically modified plants are usually more resistant and can take over other genetic lines. It happens with corn all of the time. You might have a high THC content, but you will lose the other things such as taste and would have less variation in highs. We also do not know how genetically engineered weed will affect our body when consumed. DNA is extremely complicated, we do not know what will happen when one gene is modified. It might produce a certain affect such as bigger yields, but it also might contribute the plant making differnent compounds that could be high carcinogenic. Changing one thing in DNA can contribute to multiple changes things in the plant, you might not even end up with pot at the end of the day. You might just create tobacco with THC, have fun smoking that. Have you had genetically engineered food, it taste like crap. Nature has done a wonderful job creating life, lets let it do its thing.

Many people also do not understand how genetic engineering is done. Basically you take a certain gene put it into a virus, since viruses are able to modify DNA, take that gene/virus insert it into the foreign DNA, not all the DNA will contain the new gene will contain it. So you kill all of the DNA that does not have the new gene with drugs. I know this is not very detailed but I do not feel like really explaining the process. In short genetic engineering can lead to certain viruses becoming resistant to drugs. Genetic engineering needs at least 50 more years before it should have used on the ridiculous industrial level it is now. Every one should go on Hulu and watch 'The Future of Food', it will blow your mind.

Instead we could use the tools to look at what genes contribute to certain aspects, taste, THC %, etc... so we can no what plants would be best to breed to get the results that we want. Without the guessing game of what male should be breed with what female. You could pick males and females to breeding because their DNA contained the code of blueberry taste, high yield, etc... I would love to work in such a industry, good thing I already have the skills

I say go for it, but use caution. What the best result would be to make chemical specific bud. Like to make the Thc customized, and lessen any toxins, or take the tired out would be cool. It would be cool as a medical delivery system for other meds, find a way to grow them into the bud, Maybe even future chemo bud. Its an out there thought but there is so much more to pot. So much, and since it doesnt hurt you, or the env, and isn't terribly hard to quit. . . well why is it still not federally legal, and UN says no to it too. I say go with it worldwide, i know americans can make the best smoke, and we like exotics to. Sounds like a taxible import export business, and tons more frieght and international business for airlines. Hmmmmmm. So it is the answer!!!!!

HarvestFest2010 said:

I say go for it, but use caution. What the best result would be to make chemical specific bud. Like to make the Thc customized, and lessen any toxins, or take the tired out would be cool. It would be cool as a medical delivery system for other meds, find a way to grow them into the bud, Maybe even future chemo bud. Its an out there thought but there is so much more to pot. So much, and since it doesnt hurt you, or the env, and isn't terribly hard to quit. . . well why is it still not federally legal, and UN says no to it too. I say go with it worldwide, i know americans can make the best smoke, and we like exotics to. Sounds like a taxible import export business, and tons more frieght and international business for airlines. Hmmmmmm. So it is the answer!!!!!

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The thing is that would be nice, but scientist are still not at the point where they could modify the genome of an advanced organism and it would only have the effect that they want. You could end up destroying many different strains while just trying to create one. Most of the scientist that work on genetic engineering think we should stop doing it on the industrial level. Only do it in a very closed and controlled environment. It is a VERY dangerous science that needs decades before it should introduced on such a level. I am not willing to risk the ecosystem on bud. Its not worth rushing into, you can still consume THC by eating it, that genetic engineering really isn't worth using at its present level. We are actually doing more harm then good by using it the way that we have used it. I also think it is impossible to completely take out everything that is bad for us, since many of those things are good for the plants. Plants are more advanced then many people think.

Fascinating read and very well explained. I don't know how I feel about frankenweed but then again...

For instance, imagine a forest fire where the smoke has enough THC to get every man woman and child in an adjacent city stoned.

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I think I love you Sam!

dontexist21 said:

The thing is that would be nice, but scientist are still not at the point where they could modify the genome of an advanced organism and it would only have the effect that they want. You could end up destroying many different strains while just trying to create one. Most of the scientist that work on genetic engineering think we should stop doing it on the industrial level. Only do it in a very closed and controlled environment. It is a VERY dangerous science that needs decades before it should introduced on such a level. I am not willing to risk the ecosystem on bud. Its not worth rushing into, you can still consume THC by eating it, that genetic engineering really isn't worth using at its present level. We are actually doing more harm then good by using it the way that we have used it. I also think it is impossible to completely take out everything that is bad for us, since many of those things are good for the plants. Plants are more advanced then many people think.

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Yeah, it really is kind of scary. Even with very controlled experiments.

Ever seen a black squirrel? Ask anyone in Princeton, NJ about the black squirrels that are now fairly common in town.

northernheights said:

Hi everyone,

Hope you will forgive introducing myself in this part of the forum. I just wanted you all to be aware of this:

The Biotechnology of Cannabis Sativa

It's a free discourse on the ways biotechnology (genetic engineering) may be applied to increase cannabis potency and to possibly create a whole mess of novel structural phenotypes.

Most (if not all) of the genes in the THC production pathways have been sequenced and can be obtained for free on sites like NCBI's entrez.

Sam Zwenger, the author of the pdf I linked even goes on to document actual transgenic GFP (green fluorescent protein) cannabis. Doesn't look like much, but this is a milestone in DIY genetic engineering. Next year, I hope to duplicate his results and possibly start my own transgenic cannabis projects.

Click to expand...

No offense but I really hope you don't, any genetic engineering should be done by someone that is trained and fully understands it. Messing with the DNA of any living thing should not be done unless you have a background with biochemistry, and I am not just talking about high school biology, I mean read multiple books and papers on biology and chemistry, have put in the lab work learning the techniques, and have understand the risk associated with it. You will also need to acquire many of the tools which they use to even come close to being able to duplicate it. Just the grow chamber that he uses is a extremely expensive instrument. I would not be surprised if it cost him a few thousand. In my old lab we had one that was a few years old, work well but my prof said it was just to expensive to buy another one. This guy has funding from the Clinton foundation. Also bacteria lines are really expensive, you also have to store them in a -80C freezer, costing tens of thousands, with liquid nitrogen or you cannot use them for more then 24 hrs. And creating enough bacteria to run a series of experiments can take a few days.

If you are truly interested in this I do suggest you put in the time understanding the science and ask around if you can work in a lab if you have the qualifications to learn the skills. I have been working in a lab for 2 years, and I did some genetic engineering and organic chemistry, and I know that I would not be able to duplicate his experiments properly. Not just because I still need to learn more, also because I do not have the funds to even dream about doing this.

dontexist21 said:

As a person who majors in biochemistry and has done genetic engineering on bacteria in the lab, I feel that it should NEVER be applied to any thing that we consume. Not only is there not enough information on how modifying the genes of an organism that is introduced into the wild could have harmful effects on the environment. There is all ready signs of how genetic engineering of certain types of plants such as corn have actually contributed to the reduction of different varieties of corn. I rather the THC content of weed be determined by breeders NOT by scientist.

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First of all, let me thank you for taking the time to write your ideas on this issue. I agree with you wholeheartedly that the initial forays into genetic engineering by consolidated agricultural giants, such as Monsanto, was misguided and motivated by a negative, profit-seeking outlook. I also agree with your concerns regarding unwanted gene transmission, or gene pollution.

However, I do not believe that future genetic engineering of Cannabis will result in a reduction of the variety of strains (or the gross phenotypes distinguished by differences in flavor, smell, pharmacological effects etc.).

Remember that breeding, or artificial selection, is a form of genetic engineering. The proliferation of the hundreds of distinct strains we enjoy today is the result of decades of work by committed individuals, literally traveling the world searching for breeding lines with new and exciting characteristics. I have even read reports of a stagnation of genetic diversity since this initial period of experimentation as the much of the world's commercially obtainable cannabis seeds are consolidated among a few, albeit successful seed banks.

Developments such as woody cannabis (literally Cannabis trees) are still a very long way off. For now, we can only hope to provide evidence for the possibility of using biotechnology to make better Cannabis. I have faith that the creativity and positive energy surrounding this plant will enforce an attitude of responsible and careful experimentation.

Since Cannabis is certainly not grown in the same fashion as corn or other staple crops (namely monocultured and grown in bulk from seed, using lots of pest- and herbicides), we are lucky to avoid many of the downfalls of GMO food crops right off the bat. Cannabis growers and consumers love variety and often no two people can agree on what the optimum strain should grow, look or smoke like.

It is for these reasons that I believe that the application of modern biotechnology to Cannabis will be done a way that is harmonious with both nature and ourselves.

northernheights said:

First of all, let me thank you for taking the time to write your ideas on this issue. I agree with you wholeheartedly that the initial forays into genetic engineering by consolidated agricultural giants, such as Monsanto, was misguided and motivated by a negative, profit-seeking outlook. I also agree with your concerns regarding unwanted gene transmission, or gene pollution.

However, I do not believe that future genetic engineering of Cannabis will result in a reduction of the variety of strains (or the gross phenotypes distinguished by differences in flavor, smell, pharmacological effects etc.).

Remember that breeding, or artificial selection, is a form of genetic engineering. The proliferation of the hundreds of distinct strains we enjoy today is the result of decades of work by committed individuals, literally traveling the world searching for breeding lines with new and exciting characteristics. I have even read reports of a stagnation of genetic diversity since this initial period of experimentation as the much of the world's commercially obtainable cannabis seeds are consolidated among a few, albeit successful seed banks.

Developments such as woody cannabis (literally Cannabis trees) are still a very long way off. For now, we can only hope to provide evidence for the possibility of using biotechnology to make better Cannabis. I have faith that the creativity and positive energy surrounding this plant will enforce an attitude of responsible and careful experimentation.

Since Cannabis is certainly not grown in the same fashion as corn or other staple crops (namely monocultured and grown in bulk from seed, using lots of pest- and herbicides), we are lucky to avoid many of the downfalls of GMO food crops right off the bat. Cannabis growers and consumers love variety and often no two people can agree on what the optimum strain should grow, look or smoke like.

It is for these reasons that I believe that the application of modern biotechnology to Cannabis will be done a way that is harmonious to both nature and ourselves.

Click to expand...

There is still a big difference between selecting two different males and females because they have certain traits, and changing their genetic code. The former is requires you to change the DNA code of a living organism, and by doing that you can never be certain of what will actually occur. If cannabis were to be legalized you can be sure that the same methods of cultivation that are used with corn today will be used with cannabis. I am not completely against genetic engineering, I just think that its wide spread use in the industrial sector should be stopped till we have a understanding of it. Today scientist barely understand the genetic code, so how can scientist believe that they can change the code for any living species without having harmful consequences. We are decades away from even coming close to the dream of doing this in a responsible fashion. One of the main reasons that few seed banks contain a majority of the strains is related the fact that it is still considered a illegal substance in much of the world. I believe with just legalizing it would allow different strains to cross country borders. We do not need to rush into genetic engineering, cannabis while a wonderful plant, is not worth using a science that is barely understood and still needs decades before we can properly use it in a responsible manner.

Haven't any of u seen Attack of the Killer Tomatoes. Holy crap don't mess with plant genes! The last thing we need are huge 100 pound pot plants running amuck......Whoa wait a sec I rescind that comment.

dontexist21 said:

There is still a big difference between selecting two different males and females because they have certain traits, and changing their genetic code. The former is requires you to change the DNA code of a living organism, and by doing that you can never be certain of what will actually occur. If cannabis were to be legalized you can be sure that the same methods of cultivation that are used with corn today will be used with cannabis. I am not completely against genetic engineering, I just think that its wide spread use in the industrial sector should be stopped till we have a understanding of it. Today scientist barely understand the genetic code, so how can scientist believe that they can change the code for any living species without having harmful consequences. We are decades away from even coming close to the dream of doing this in a responsible fashion. One of the main reasons that few seed banks contain a majority of the strains is related the fact that it is still considered a illegal substance in much of the world. I believe with just legalizing it would allow different strains to cross country borders. We do not need to rush into genetic engineering, cannabis while a wonderful plant, is not worth using a science that is barely understood and still needs decades before we can properly use it in a responsible manner.

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Take for example, the developing medical marijuana distribution system in California. Increasingly, trends have shifted towards protecting the health of the consumer at all costs.

This attitude of responsibility involves complicated tests and lab equipment that are very costly. Yet the purveyors of these new safety standards know that for medicine to be effective, it must be delivered with as little harmful side effects and outside contamination as possible.

[url said:

http://www.cannabisnews.org/united-states-cannabis-news/medicalmarijuana/the-manhattan-project-of-marijuana/][/url] The East Bay’s first pot lab looks like a bachelor pad with a locked room in the back. The building is of recent construction with high ceilings and stained carpets, mismatched furniture, and a congenial guard dog, belonging to Addison.

It’s a little cooler in the locked back room. The place hums like the inside of a busy copy store. The lab’s centerpiece — the gas chromatograph — squats on a work bench in the back studded with yellow samples in a carousel feeding into an auto-sampler. Inside the device, a flame ion detector and mass spectrometer offer two different snapshots of the prepared samples. Underneath, an $80,000 hydrogen generator hums a steady supply into the chromatograph. Tanks of oxygen and air also feed the device. Off to one side, a monitor flicks line graphs. Books from Agilent Tech, Sigma Life Sciences, and Aldrich Chemistry line the bookshelf.

Dave runs through the process of documenting and preparing the sample. The gas chromatograph needs just a microliter-size sample to test; less than a rain drop. So the lab’s main methodology turns the sample packets of green bud into a diluted liquid extraction. First, the lab tech does the paperwork, and dons gloves and gear. Addison chops up a half-gram under a sterile hood and places the sample in a vial, then adds a controlled amount of Hexane — a special-use solvent.

The mix goes into a sonicator, an ultra-sonic jeweler’s tool. It vibrates at a high enough frequency to rupture the cell membranes of the plant. The liquid is then diluted to just hundredths of a percent and an extraction is loaded into a little test vial.

Rows and rows of vials are then fed into the gas chromatograph on a timer. Inside the machine it’s like CSI — but for ganja.

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Pics of their beautiful operation can be found here:
http://www.harborsidehealthcenter.com/gallerypages/index.html

northernheights said:

Take for example, the developing medical marijuana distribution system in California. Increasingly, trends have shifted towards protecting the health of the consumer at all costs.

This attitude of responsibility involves complicated tests and lab equipment that are very costly. Yet the purveyors of these new safety standards know that for medicine to be effective, it must be delivered with as little harmful side effects and outside contamination as possible.




Pics of their beautiful operation can be found here:
http://www.harborsidehealthcenter.com/gallerypages/index.html

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My main point is genetic engineering as a whole is something that is barely understood. We do not understand how DNA functions, even when dealing with lower level organism such as plants. How can we modify the DNA when we don't understand it. There is no responsible way of using genetic engineering in its current form. A few decades when we have done much more work on it I would like to see it being used on a wider scale, but not now, not when it would do more harm then good. Even when people with the best intentions cannot use it responsibly in its current state. I also do not think that we need to. There is a reason plants produce harmful chemicals, it helps them survive, you might be doing more harm then good. And THC and many of the other helpful compounds can always be extracted using simple procedures. Hell if you gave me a few simple widely used solvents, a few pieces of lab equipment, I could give you some of the purest extract that you have ever had. Using barely any heat, so no lost of THC and no chance of solvents being left in the final product. And with time the separate compounds can be separated if I used some other methods, this would mean you would only get the specific pain relief that the patient needs. And no need to be using untested genetic engineering.