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Genome Has Been Sequenced

GreenintheThumb

fuck the ticket, bought the ride
Veteran
August 18, 2011 10:00 AM Eastern Daylight Time
Medicinal Genomics Sequences the Cannabis Genome to Assemble the Largest Known Gene Collection of this Therapeutic Plant

The data, representing a 65,000 fold increase in what is publicly known about the genome, is being made readily available to researchers and scientists

MARBLEHEAD, Mass. and AMSTERDAM--(BUSINESS WIRE)--Medicinal Genomics, a pioneer in the genomics of medicinal plants, today announced that the company has sequenced the entire genome of Cannabis sativa and Cannabis indica, assembling the largest known gene collection of this therapeutic plant. The DNA purification was performed at the company’s research facility in Amsterdam, and the sequencing was conducted by several service providers, including the latest long read technology from Roche’s 454 sequencing center.

“Despite compelling evidence of the therapeutic benefits of Cannabis, very little genomics research has been performed in this area”

Previously, only two million bases of Cannabis sequence have been deposited in GenBank, a sequence database provided by the National Center for Biotechnology Information (NCBI). To date, Medicinal Genomics has privately sequenced over 131 billion bases of sequence, which represents a 65,000 fold increase in what has been publicly shared about the Cannabis genome. Concurrently, Medicinal Genomics has published the raw reads from Cannabis sativa on Amazon’s EC2, a public cloud computing service, giving the scientific community access to conduct further research. The Cannabis indica genome sequence will be made available on the EC2 in a few weeks as well. The genome annotations will be made accessible via an iPad application that the company expects to launch in the fall.

“Despite compelling evidence of the therapeutic benefits of Cannabis, very little genomics research has been performed in this area,” said Kevin McKernan, founder and head of scientific operations of Medicinal Genomics. “Cannabis was one of the most difficult genomes that I ever sequenced, and even though only a draft assembly has been constructed, it is important to provide the scientific community with the raw data as quickly as possible. Ongoing scientific research suggests that some of the non-toxic compounds in this plant may ultimately prove to be powerful therapeutics that can treat a wide range of health conditions, including cancer and inflammatory diseases.”

More than 40 U.S. Food and Drug Administration (FDA) approved clinical trials evaluating Cannabis as a therapy have been completed or are underway, according to information obtained from www.clinicaltrials.gov.

“This is a significant accomplishment,” said Richard Gibbs, Ph.D., director of the human genome sequencing center at the Baylor College of Medicine. “It is excellent to see rapid data release policies being upheld by public and private organizations, particularly when it comes to such challenging genomes.”

With the complete genome in hand, researchers can begin to identify non-psychoactive compounds or enzyme pathways to better elucidate the therapeutic benefits of Cannabis, including the plant’s anti-cancer properties. These pathways can be optimized in the plant or cloned into other hosts for more efficient biologic production. In addition, it may be possible through genome directed breeding to attenuate the psychoactive effects of Cannabis, while enhancing the medicinal aspects.

Medicinal Benefits of Cannabis

Ongoing scientific research suggests that the Cannabis plant harbors beneficial compounds as it pertains to cancer apoptosis, antiemesis for HIV and chemotherapy patients, reduction of muscle spasms for multiple sclerosis patients, as well as the treatment of glaucoma, inflammatory diseases, anxiety, and post-traumatic stress disorder, among other health conditions. Of the 85 identified Cannabinoids in the plant, only one is independently psychoactive. The other 84 appear to be non-psychoactive and are still powerful therapeutics. Many Cannabinoids have been bred to low expression levels over the years but are excellent small molecule therapeutic drug candidates for cancer and inflammatory diseases. The genome sequence can be utilized to design breeding strategies to resurrect these nearly extinct pathways. In addition, Cannabinoids have also been shown to have a very favorable therapeutic index making them a unique class of anti-cancer compounds because the lethal dose is much higher than the effective dose. Furthermore, Cannabinoids are less addictive analgesics than opiates, and target different pain pathways in the body.

Another benefit of Cannabis as a source for a therapeutic drug is that its toxicity is low and well-known. This is unique in drug discovery where every drug candidate must go through expensive and lengthy clinical trials to establish its safety. With Cannabis, side effects have previously been established and might be mitigated with genome directed breeding, whereas many drug candidates are not easily modified and end up failing in clinical trials because of adverse effects on a small percentage of the population.

The American Medical Association, as well as the American College of Physicians each have independently issued reports on the medicinal benefits of Cannabis to justify a reclassification of the plant so it can be more easily researched. By digitizing the genome of Cannabis, researchers from around the world can now study this plant without having to handle it. Assembling the complete Cannabis genome was a logical first step to enable more comprehensive scientific research to begin.

About Medicinal Genomics

Medicinal Genomics is the first known organization to sequence the complete genomes of Cannabis sativa and Cannabis indica. Aside from being a useful tool to discover small molecule therapeutic drug candidates, there are many other commercial applications of these sequences, including the design of genomic assays that measure genes predictive of Cannabinoid levels in the plants at seedling stages. Use of this information will enhance breeding strategies to develop high CBD or other medicinal traits into strains. These genomic assays can also be utilized to help regulate the medicinal Cannabis market through better oversight and labeling. The non psycho-active plant known as hemp is also a productive plant for fiber production and may benefit from genome sequencing in its pursuit to improve breeding programs.

The company was founded by Kevin McKernan, a recognized thought leader and innovator in DNA sequencing. Kevin co-invented a magnetic bead-based chemistry, called SPRI, which enabled fully automated nucleic acid purification for the Human Genome Project. Based on the SPRI technology, Kevin, along with his two brothers, founded Agencourt Bioscience, an efficient, low cost sequencing laboratory. Agencourt was the only commercial laboratory selected to participate in the Human, Rat, and Puffer Fish Genome Projects, in addition to serving as the lone sequencing facility for the Mammalian Gene Collection. Agencourt was sold to Beckman Coulter in 2005. Prior to the sale to Beckman, Agencourt spun off its subsidiary, Agencourt Personal Genomics, to its shareholders. Agencourt Personal Genomics was a next generation sequencing (NGS) project that Kevin had also co-invented, which subsequently became known as SOLiD. Agencourt Personal Genomics created an instrument that advanced the accuracy and throughput of NGS, lowering the cost of sequencing by a factor of 100,000. Nine months after the spin out, Agencourt Personal Genomics was purchased by Applied Biosystems, the then leader in DNA sequencers. Kevin remained with Applied Biosystems for 5 years, even after Applied Biosystems was acquired by Life Technologies, to complete the family of NGS products. He most recently served as vice president of advanced research and head of research and development for the Ion Torrent division of Life Technologies.

Medicinal Genomics maintains its corporate headquarters in Marblehead, Mass. All scientific operations are conducted at the company’s research facility in Amsterdam, Netherlands. For further information, please visit www.medicinalgenomics.com.
 

Chimera

Genetic Resource Management
Veteran
It was Chemdawg they sequenced.

Then at least we know the intersex sequence(s) are in there somewhere!

Seriously though, other reports state the DNA was extracted from single clones of LA Confidential and Chocolope.

None of that really matters, it's not a full sequence only a partial draft, but it does give us some serious information to use as breeders.

The real question is: How many breeders on ICMAG or other seed selling sites know how to use the information? ;)

Truly an amazing and important step for cannabis breeding, selection and improvement. I for one am very thankful for this company making the information accessible to all... we are really about to make some serious steps ahead.

A momentous day for Cannabis, hurrah!

-Chimera
 

zymos

Jammin'!
Veteran
I can't tell from the press release- have they really sequenced the genome, or just a bigger chunk of it than had been done before?
 

spurr

Active member
Veteran
Arrg! Another thread pulling me out of social groups only! Damn you GitT and Chimera! Both of you are on my shit list, with mofeta :D

chimera said:
The real question is: How many breeders on ICMAG or other seed selling sites know how to use the information?

While I know (a decent) amount about DNA marker assisted breeding, what I know pales to what you and folks like mofeta know. However, I can humbly suggest I know 'more than the average bear' ... and we all (yous' know who I'm writing about ;)) need to get our shit together ... time for a take over!

(from my Blockhead Society social group)

Note: this is not a process of genetic engineering (GE), where by definition one ends up with a genetically modified organism (GMO), i.e., a transgenic plant.

As an example, I will be looking at DNA makers to see if the strain (i.e., plant) I am working with is male [1], cannabinoids profile, chemotype, etc.

Using DNA marker assisted breeding is like giving a traditional breeder x-ray eyes, so they can look at DNA code, and nothing more in terms of GE or GMO. DNA marker assisted breeding is a way to augment and enhance selective (traditional) breeding. However, I think using such breeding methods for Cannabis spp. are in their infancy.
[1] sexual differentiation of Cannabis spp. occurs when 4th internode leaf set appears.​
Please see these web pages and videos from the National Science Museum project: "Secret of plant genomes revealed!"
(good intro into DNA markers, PCR machines, etc.)
- Video #3 on genomics (not GE or making GMO)
http://www.plantgenomesecrets.org/w...nome?width=640&height=425#video_container_101


- Whole video on genomics (incl. GE and GMO):
http://www.plantgenomesecrets.org/w...show?width=640&height=425#video_container_134


- Genomics slide show (using PCR machine, etc.):
http://www.plantgenomesecrets.org/genome-basics


- Traditional breeding. Maker assisted. Transgenic plants.
http://www.plantgenomesecrets.org/g...ng-marker-assisted-breeding-transgenic-plants


How do you change a plant? Well—as the video to the left shows—humans have been doing it since the beginning of civilization through a process called selective breeding. When a farmer finds a plant with desirable traits—like bigger fruit, leafier greens, or disease resistance—that plant is bred with another plant. If the farmer is lucky, that trait is passed on to the baby plants. But, it is a long, hit and miss process that can go on for generations before the desired quality becomes consistent.

Marker assisted breeding is very similar to selective breeding, but it uses tools called genetic markers to figure out which plants would make the best parents. Genetic markers reveal the exact genes responsible for a particular trait. This lets breeders locate parents that are guaranteed to reproduce the qualities they want. Taking the guesswork out of selective breeding can save months or years in the development of an improved crop.

Genetic markers are genomic tools, but the plants produced using them are not "genetically engineered". Their produce cannot be labeled as “genetically engineered”. Genetic engineering, on the other hand, allows scientists to transfer DNA from one organism to another—even between different species. These plants are called transgenic plants [a type of GMO].

The sequencing of the genome will create a base of knowledge—a “road map”—for scientists as they discover the relationships between genes and how they work. The more scientists learn about how nature develops new plant species, the better that knowledge can be applied to building better plants.

Misc. relevant Cannabis spp. studies:
Genetics and Marker-assisted Selection of the Chemotype in Cannabis sativa L.
Daniela Pacifico, Francesca Miselli, Mirta Micheler, Andrea Carboni and Paolo Ranalli, et al.
Molecular Breeding, 2006, Volume 17, Number 3, Pages 257-268


The Inheritance of Chemical Phenotype in Cannabis sativa L.
Etienne P. M. de Meijer, Manuela Bagatta, Andrea Carboni, Paola Crucitti, V. M. Cristiana Moliterni, Paolo Ranalli, and Giuseppe Mandolino
Genetics, Vol. 163, 335-346, January 2003


The Applications of Molecular Markers in Genetics and Breeding of Hemp
Giuseppe Mandolino and Paolo Ranalli
Journal of Industrial Hemp, Volume 7, Issue 1, 2002
DOI: 10.1300/J237v07n01_03


Potential of marker-assisted selection in hemp genetic improvement
Giuseppe Mandolino, Andrea Carboni
Euphytica, Volume 140, Numbers 1-2, 107-120 (2004)
DOI: 10.1007/s10681-004-4759-6


Current status and future scenarios of hemp breeding
Paolo Ranalli
Euphytica, Volume 140, Numbers 1-2, 121-131 (2004)
DOI: 10.1007/s10681-004-4760-0


Marker assisted selection and genomics of industrial plants
Giuseppe Mandolino
Improvement of Crop Plants for Industrial End Uses, 2007, 59-82
DOI: 10.1007/978-1-4020-5486-0_3


Advances of hemp sector in Italy and its perspectives
P. RANALLI
Istituto Sperimentale per le Colture Industriali, Via di Corticella 133, 40128 Bologna


Development of microsatellite markers in Cannabis sativa for DNA typing and genetic relatedness analyses
H. J. Alghanim and J. R. Almirall
Analytical and Bioanalytical Chemistry, Volume 376, Number 8, 1225-1233, (2003)
DOI: 10.1007/s00216-003-1984-0


Genetic individualization of Cannabis sativa by a short tandem repeat multiplex system
Maria A. Mendoza, DeEtta K. Mills, Hemant Lata, Suman Chandra, Mahmoud A. ElSohly and Jose R. Almirall
Analytical and Bioanalytical Chemistry, Volume 393, Number 2, 719-726, (2009)
DOI: 10.1007/s00216-008-2500-3


Inter simple sequence repeats separate efficiently hemp from marijuana (Cannabis sativa L.)
Erdogan E. Hakki, Seyit A. Kayis, Emine Pinarkara, Ayla Sag
Electronic Journal of Biotechnology, Vol. 10 No. 4, Issue of October 15, (2007)
ISSN: 0717-3458


Secondary metabolism in cannabis
Isvett Josefina Flores-Sanchez and Robert Verpoorte
Phytochemistry Reviews, Volume 7, Number 3, 615-639, (2008)
DOI: 10.1007/s11101-008-9094-4
Sex relevant C. spp. studies:
Boys and Girls Come Out to Play: The Molecular Biology of Dioecious Plants
Charles Ainsworth
Annals of Botany 86 : 211-221, 2000
doi:10.1006/anbo.2000.1201


Identification of DNA markers linked to the male sex in dioecious hemp (Cannabis sativa L.)
G. Mandolino, A. Carboni, S. Forapani, V. Faeti and P. Ranalli
TAG Theoretical and Applied Genetics, Volume 98, Number 1, 86-92 (1999)
DOI: 10.1007/s001220051043


Sex-linked AFLP markers indicate a pseudoautosomal region in hemp (Cannabis sativa L.)
A. Peil, H. Flachowsky, E. Schumann and W. E. Weber
TAG Theoretical and Applied Genetics, Volume 107, Number 1, 102-109,
DOI: 10.1007/s00122-003-1212-5


Occurrence and frequency of putatively Y chromosome linked DNA markers in Cannabis sativa L.
Giuseppe Mandolino, Andrea Carboni, Manuela Bagatta, V.M. Cristiana Moliterni and Paolo Ranalli
Euphytica, 2002, Volume 126, Number 2, Pages 211-218


Novel male-specific molecular markers (MADC5, MADC6) in hemp
Ottó Törjék, Nándor Bucherna, Erzsébet Kiss, Hajnalka Homoki, Zsuzsanna Finta-Korpelová, Iván Bócsa, István Nagy and László E. Heszky
Euphytica, Volume 127, Number 2, 209-218, DOI: 10.1023/A:1020204729122


The sexual differentiation of Cannabis sativa L.: A morphological and molecular study
V. M. Cristiana Moliterni, Luigi Cattivelli, P. Ranalli and Giuseppe Mandolino
Euphytica, 2004, Volume 140, Numbers 1-2, Pages 95-106
Here are some links and info about work with C. spp. genome and genes coding cannibanolic acid expression, etc. Does anyone have additional resources or data they could add?

http://www.plantgdb.org/search/misc/plantlistconstruction.php?mySpecies=Cannabis sativa

http://www.ncbi.nlm.nih.gov/nuccore/AB292684.1

http://www.ncbi.nlm.nih.gov/nuccore?term="Cannabis"[Organism]&cmd=DetailsSearch (copy the whole URL from "http" to "DetailsSearch")


Now, where the hell did I put my army of PCR machines, -80 C freezer, microcentrifuge, and etc.?!

index.cfm



_80Freezer-2.jpg


sm-DNA%20in%20freezer.jpg




MicroCentrifuge.jpg
 
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spurr

Active member
Veteran
The genome annotations will be made accessible via an iPad application that the company expects to launch in the fall.

^^^ This is my only reason for wanting an iPad!
 

mofeta

Member
Veteran
This is good news!

I think this shows how one person can make a huge impact on society. If we had to wait for a government to do this work we would have had to wait for a long time.

This guy Kevin McKernan is a fucking super-genius. His inventions made the Human Genome Project (among others) possible, and made him a very wealthy man. We are lucky he decided to make this huge advance a reality.

McKernan is one of the only individuals on the planet that could do this. If he says "Cannabis was one of the most difficult genomes that I ever sequenced,.." then good luck for a normal human being! He also spent 200K of his own money to do it.

McKernan is also adhering to the "Bermuda Principles" of instant, pre-publication release of data, which he didn't have to do, as he received no government funding. This is real philanthropy.

McKernan and his brothers are super-sharp businessmen too, and they intend to make a lot of money doing this. There will be numerous patents involved. Seeing as they got where they are by inventing gene-sequencing equipment I think this sentence from the press release gives an idea of one of the things they are working on:

...there are many other commercial applications of these sequences, including the design of genomic assays that measure genes predictive of Cannabinoid levels in the plants at seedling stages.

How would you like to have that product! Wow! Males too! WOW!
 

mofeta

Member
Veteran
Let that last thought resonate in you mind:

A process that accurately predicts phenotype of both male and female seedlings.
 

GreenintheThumb

fuck the ticket, bought the ride
Veteran
Then at least we know the intersex sequence(s) are in there somewhere!

:laughing:

Seriously though, other reports state the DNA was extracted from single clones of LA Confidential and Chocolope.

None of that really matters, it's not a full sequence only a partial draft, but it does give us some serious information to use as breeders.

http://csativa.elasticbeanstalk.com/

This is where I read it was the whole genome and that it is Chemdawg

The real question is: How many breeders on ICMAG or other seed selling sites know how to use the information? ;)

Oh don't you worry brother, some of us know what to do with this data. Like...3 of us. Luckily we're all on the same team. By chance I just met with a start up founded with Google money that builds specialized computing hardware and software pretty much in competition with Nimbus. We'll see what they have to say about this as well.
 

mofeta

Member
Veteran
Aren't the MADC markers old news?

Sure.

That's not what I meant though.

I'm talking about predicting expression of other phenotypic traits like cannabinoid/terpene profile, growth habit, color, pest resistance etc.

Not having to grow out female seedlings to maturity to identify which ones are on target is would be a huge advance, but mainly a time and space saving one.

Being able to identify male seedlings that are on target for female traits normally masked by their maleness, that is huge. The current methods of deducing male genotype/potential phenotype (examining progeny from crosses, backcrosses, etc for clues) would be relegated to the trash bin of history, where it belongs. Determining which male seedlings out of 1000 are the best for your goals is at present basically a trial and error crapshoot. Hasn't every serious breeder thought when cullling males "did I just kill THE ONE?". A good genetic screening process would make it possible to identify the right males from unlimited numbers, with certainty (or as many as you could afford to screen, anyway).

Of course, these are obvious potential applications of having the genome, it's why we wanted it, no? But the thing is, I always thought that the genome would be revealed by an academic/government entity and it would be up to the (remarkably few) Cannabis breeders versed in molecular genetics to sort out the genes and their interactions that produce visible phenotype. Of course folks like GW would jump all over it, but keep it proprietary. I envisioned a long, difficult, expensive slog, led by guys like Chimera, to make the revealed genome useful to us.

That's not the way it turned out, though. McKernan's almost superhuman abilities in this field, along with his stated desire for "design of genomic assays that measure genes predictive of Cannabinoid levels in the plants at seedling stages" will make our wishful thinking reality, and sooner rather than later.

I would love to have access to a genomic assay for traits in Cannabis, and it really is unparalleled good fortune that one of the leading lights of molecular genetics has decided to bend reality to his will and make it so.
 

spurr

Active member
Veteran
Medicinal Genomics, a pioneer in the genomics of medicinal plants, today announced that the company has sequenced the entire genome of Cannabis sativa and Cannabis indica ...
Maybe that incorrect wording is the fault of the article author? Or is Medical Genomics claiming it's a fact there are two species of the genus Cannabis? As of this time, AFAIK, there is insufficient genetic proof to state as a fact the genus Cannabis is monotypic or polytypic. (I for one think it will bear out as polytypic, which is why I use C. spp. instead of C. sp.)

@ mofeta,

I agree with you 100%, it's amazing what Kevin McKernan and his team (of brothers?) accomplished. And I am very thankful he/they were kind enough to publish the data. I see for commercial use Medial Genomics is charging fees for use of data, and I hope they make lots of money, but I also hope they don't charge exorbitant fee(s).
 
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spurr

Active member
Veteran
Of the 85 identified Cannabinoids in the plant, only one is independently psychoactive. The other 84 appear to be non-psychoactive and are still powerful therapeutics.
FWIW and IIRC, Sam Skunkman wrote to me that via. personal correspondence, Dr. Mahmoud A. Elsohly (of ol' Miss, IIRC) informed Sam he had qualified over 90 cannabinoids. I'll see if I can find the post from Sam ... yup, here it is:

Sam_Skunkman said:
Dr Mahmoud A. ElSohly recently told me more then 90, I have seen publications that list 85, but I need to remember which one. Not sure if all 90 are published yet.
 

Sam_Skunkman

"RESIN BREEDER"
Moderator
Veteran
I was very happy to see the news, but no one yet knows how to use the data, but they will learn.
People always talk about MADC, but FADC are known and maybe intersexed markers will be found? That would be real interesting...
Journal of Industrial Hemp 2003 Vol 8 issue 1 page 5-9, Female-Associated DNA Polymorphisms of Hemp (Cannabis sativa L.), Hong Shao, Shu-Juan Song, Robert C. Clarke

-SamS
 

GreenintheThumb

fuck the ticket, bought the ride
Veteran
So they have 20 cultivars in line for sequencing. They already completed LA Confidential as well as Chemdawg. Chocalope they're working on as well as others.

Sam did you by chance give them any of you cannabinoid specific chemovars? Seems like a no brainer.
 

mofeta

Member
Veteran
but no one yet knows how to use the data, but they will learn.


I assume they will use the Genome-Wide Association Study (GWAS) methodology that is used in studying human diseases, mapping variation, looking for SNPs.

So they have 20 cultivars in line for sequencing.

That's a good start. The more the better. Association studies work best when you have a lot of individuals to compare.

Sam did you by chance give them any of you cannabinoid specific chemovars? Seems like a no brainer.

This is a good idea. The more different the phenotype the more the SNPS stand out. You would get more useful data comparing Chemdog to some weird CBD only chemovar than you would comparing it to something relatively similar, like LA Confidential.
 

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