In this article, Gary Yates, chief scientific officer at PharmaSeeds, draws comparisons between the central biological dogma and setting up a new cultivation site.
The licensed cannabis space is expanding at a steady rate, and as such, there is a combination of new players to the game as well as older heads which have seen the industry move forward in the recent past. One thing that becomes apparent to new entrants as they progress along the journey is the seemingly never-ending variables that influence a Licensed Producers (LPs) level of success. On paper, it can look like all you need is a good facility set-up, the right genetics and successful cultivation – three separate elements that are very co-dependent.
However, as LPs start to peel back the layers of these three basic inputs, it can become overwhelmingly complex very quickly. Although the license itself acts as the gateway to production and is therefore more of a binary component, the level of input required to attain a license is extremely detailed, time-consuming, and very costly. This is especially true in stricter regions – the UK, for example – and is further compounded by the infrastructure of the facility, which must be appropriate for all elements downstream, such as cultivation methods, as well satisfying the license requirements and any further regulatory accreditation such as GMP.
This has a likeness to one of the major biological principles which state that biological information flows from DNA to RNA to protein. This central dogma governs biological systems where the relationship between DNA and the functional molecules they give rise to – proteins – have a logical and comprehensible flow. This is so fundamental that any first-year biology undergraduate will be able to recite the order in which this information flows – DNA to RNA to Protein (Figure1). But much like the description above of licensed cannabis production, the level of complexity deepens very quickly when the layers are peeled back and expose the underlying mechanisms of action.
Biology.. easy as pi…squared
What this central biological dogma really means is that the genes in the DNA are transcribed (or converted) to a special type of RNA molecule known as mRNA (messenger RNA), and this mRNA is then translated (or converted) into a protein, A.K.A. an amino acid sequence with a complex 3D structure. The language used in biology to describe each of these suites of molecules is Genome (All DNA including genes), Transcriptome (all the DNA that is converted to RNA), and the Proteome (all products resulting from translation otherwise put, conversion from RNA to amino acid), including the subsequent modifications (Figure1).
Figure 1. The central biological dogma. This diagram shows the way information flows from DNA to protein through the mRNA intermediate. Listed in text boxes coloured blue, gold, and red are examples of modification (DNA, RNA and protein respectively) which can occur at each step and can affect the end product.
The three elements described also represent important regulation/control or checkpoints. A simplified example of the variables for each of these can be described by:
1) How much is the gene expressed (i.e. how much is turned into RNA)
2) How much of the RNA gets degraded vs that which gets translated, and
3) Of the translated protein, what amount is present at any given time
Relating back to the licensed cannabis industry and sticking with the three elements aforementioned (bearing in mind there are many ways to divide the requirements of a successful LP), facility set-up, the right genetics and successful cultivation, an example of variables for each could be:
1) When does the license begin and end
2) What is the output of the chosen genetics
3) Does the finished product meet the standards of the offtake agreement
Complexity – the fractals of reality
Of course, in reality, there are numerous other considerations for each step in both scenarios, with both quantitative and qualitative elements to each, but more so, there is a plethora of other important steps which must be included to ensure success at each of the ‘big three’ stages. For example, DNA contains genes, and genes can be active or inactive (on or off), and genes can be highly expressed or lowly expressed.
One gene can give rise to more than one version of its protein product through, for example, modification to the mRNA via a process called ‘alternative splicing’. Gene expression can be temporally regulated and/or can be controlled by master regulatory elements which orchestrate entire suites of genes – such as flowering initiation. There are several ways the mRNA can be modified, and a whole host of modifications which can be made to the protein itself (Figure 2).
Figure 2. Examples of modification which can occur to proteins. Some modifications will change protein function depending on where they sit on the protein, and how many times the modification occurs throughout the protein.
What this means is that there is significantly more than one variation downstream of the DNA than there is number of genes in the DNA. For example, an organism with 20-25,000 genes is capable of producing over 1 million proteins/protein variants (Figure 3). This perfectly captures how variable the outcome of a biological system can be, and even though not every variable is considered here, it helps conceptualise the unpredictable nature of biology through modifications, variations and combinations that influence the output.*
Figure 3. Genes to Proteins. Infographic showing the increase of complexity as information flows from DNA to protein. At each step there are numerous modifications that amplify the number of products from the previous step, resulting in a highly variable end product
In the comparison, the license can therefore be likened to whether any one cultivar has a particular trait or not, and the infrastructure itself could represent how genes are expressed. As this analogy continues, and when considering the choice of genetics and the success of the cultivation, obviously the level of complexity jumps exponentially as cultivators consider cultivation techniques, feed delivery systems and media, genetics and germplasm, personnel, and so on – all of which can also be further subcategorised.
Having the correct set up is crucial to productivity and therefore when a license holder is seeking their genetic starting material it must be in line with the setup and the desired output. And even after a successful grow, there are considerations regarding product readiness such as harvest method and trimming, drying and curing, packaging, storage and so on – comparisons with the modification that can be added to proteins (figure 2)
These steps are in every way as essential as any prior step, as the condition of the final product can still be vetoed by the buyer if it does not meet their required standard.
The reason these two seemingly unrelated topics have parallels stems from the numerous variables involved in each scenario. With the biology, quantifying is made easier as there are decades of research to prop up the numbers, as well as the ever-growing layers of regulation and control which are frequently being discovered and written about. This, along with subjects being divided into new disciplines, creates a constant stream of new information which is being built upon from the preceding data and published work.
However, in the licensed cannabis space, the variables are much harder to quantify and there is not the same level of consistent data to fall back on – in fact, on many occasions, licensed producers are so busy dealing with one set of variables that they often overlook another set of variables. This, like the science counterpart, is better remedied by compartmentalisation of each discipline. Supply of genetics, for example, is best served with a professional growing plan and cultivation instructions to optimise the genetic potential. This will only work in facilities where the setup is suitable for the recommended genetics, hence the need to have specialist suppliers.
By having these specialists focused on their area of expertise, i.e. matching genetics to the environment they are intended to be grown in, and by having the entire system orchestrated from build of site to offtake, there is then a much greater likelihood that everything functions as it should. If a gene should pick up a mutation that affects the protein product of that gene, then there is a good chance the function of the protein (and therefore function of the gene) could be altered, resulting in disease.
This is similar to a part of a growing facility’s infrastructure failing and causing unfavourable conditions which result in a drop of yield or quality in the plant. Much like in biological systems, small changes at the input stage (genes) can cause devastating changes to the ability of that biological system to function properly.
This is also true in modern cannabis cultivation where the lack of real-time monitoring, coupled with the lack of good contingencies can further escalate those which would otherwise be small problems when they arise. As with human health, where early detection of disease often stands the patient in better stead for treatment and recovery, so does early identification of problems in a grow chamber.
If an issue can be known to the grower before it has a chance to affect the plants then there is obviously less chance of it manifesting into a real problem, especially if good contingencies are in place. Selection of the correct infrastructure, genetics, growing system and personnel can all help reduce the risk to crops.
When the genetic information flow was first published and biologists started to understand DNA, RNA and protein function, no one could have predicted the numerous levels of regulation and other influential factors involved in controlling these biological systems. This is echoed in the license cannabis industry where the further into the process anyone goes, the more learning must be done. Much like in biology there is too much information throughout all the disciplines for one person to truly master it all, so as with any industry, dedicated personnel controlling finance, sales, cultivation and so on are needed in order to maximise the return on investment and fulfil the industry’s potential.
*During my undergraduate studies, the human genetics lecturer described the maths involved in multifactorial inheritance (many genes involved in determining the phenotype, for example predisposition to a disease state such as heart disease) as surpassing the complexity of maths use in rocket science calculations – an interesting and surprising little fact.
Chief scientific officer
New research could help cultivators control THC and CBD levels in crops
A team of researchers has used firefly genes to understand cannabis biology.
A better understanding of how cannabis produces THC means scientists could selectively knock out the enzyme that synthesises THC using genome editing techniques such as CRISPR. This would produce plants with lower levels of, or no levels of, THC.
With strict regulations surrounding the levels of CBD and THC in cultivated cannabis, controlling these levels is vital to prevent destruction of crops and lost licences, for example.
Cannabinoids are produced by trichomes, the small, spikey and sticky protrusions on the surface of cannabis flowers, however, scientists know very little about how cannabinoid biosynthesis is controlled.
To discover the underlying molecular mechanisms behind trichrome development and cannabinoid synthesis, Yi Ma, research assistant professor, and Gerry Berkowitz, professor in UConn’s College of Agriculture, Health and Natural Resources received funding through the National Research Initiative from the US Department of Agriculture.
The research has been published in the journal Plants.
Berkowitz and Ma, and former graduate students Samuel Haiden and Peter Apicella, have discovered transcription factors responsible for trichome initiation and cannabinoid biosynthesis.
Transcription factors are molecules that determine if a piece of an organism’s DNA will be transcribed into RNA, and thus expressed. In this case, the transcription factors cause epidermal cells on the flowers to morph into trichomes.
With this new grant, the researchers will continue to explore how these transcription factors play a role in trichome development during flower maturation.
Berkowitz and Ma will clone the promoters – the part of DNA that transcription factors bind to – of interest, and will then put the promoters into the cells of a model plant along with a copy of the gene that makes fireflies light up, known as firefly luciferase; the luciferase is fused to the cannabis promoter so if the promoter is activated by a signal, the luciferase reporter will generate light.
Berkowitz commented: “It’s a nifty way to evaluate signals that orchestrate cannabinoid synthesis and trichome development.”
The researchers will load the cloned promoters and luciferase into a plasmid. Plasmids are circular DNA molecules that can replicate independently of the chromosomes. This allows the scientists to express the genes of interest even though they aren’t part of the plant’s genomic DNA. They will deliver these plasmids into the plant leaves or protoplasts, plant cells without the cell wall.
When the promoter controlling luciferase expression comes into contact with the transcription factors responsible for trichome development (or triggered by other signals such as plant hormones), the luciferase ‘reporter’ will produce light.
Ma and Berkowitz will use an instrument called a luminometer, which measures how much light comes from the sample. This will tell the researchers if the promoter regions they are looking at are controlled by transcription factors responsible for increasing trichome development or modulating genes that code for cannabinoid biosynthetic enzymes. They can also learn if the promoters respond to hormonal signals.
In prior work underlying the rationale for this experimental approach, Ma and Berkowitz along with graduate student Peter Apicella found that the enzyme that makes THC in cannabis trichomes may not be the critical limiting step regulating THC production, but rather the generation of the precursor for THC (and CBD) production and the transporter-facilitated shuttling of the precursor to the extracellular bulb might be key determinants in developing cannabis strains with high THC or CBD.
Most cannabis farmers grow hemp, a variety of cannabis with naturally lower THC levels than marijuana. Currently, most hemp varieties that have high CBD levels also contain unacceptably high levels of THC. This is likely because the hemp plants still make the enzyme that produces THC. If the plant contains over 0.3% THC, it is considered federally illegal and, in many cases, must be destroyed.
The researchers said: “We envision that the fundamental knowledge obtained can be translated into novel genetic tools and strategies to improve the cannabinoid profile, aid hemp farmers with the common problem of overproducing THC, and benefit human health.”
This knowledge could lead to the production of cannabis plants that produce more of a desired cannabinoid, making it more valuable and profitable.
As well as sharing these findings with cannabis scientists, industry, and growers, the researchers will incorporate this new knowledge into UConn courses on cannabis horticulture.
This grant will also support the training of undergraduates interested in cannabis research, providing them with the skills to enter the workforce.
Planning application submitted for Isle of Man cannabis facility
The facility will include a science and innovation centre.
Plans to kickstart the medical cannabis industry on the Isle of Man have progressed with the submission of a formal planning application for a cultivation facility on the island.
Earlier this year, Peel NRE proposed to establish a multi-million-pound science innovation and research centre (SIRC), sustainable energy park and medical cannabis facility on the Isle of Man.
The regeneration and clean energy specialist has now applied for planning consent for its proposals for the 72-acre site in between the A6 and A5 on New Castletown Road.
The plan, which represents an investment of over £150m from Peel NRE, would create around 250 jobs across a range of skills from botany and technology to security and exports. More than 178,000 sqft of cannabis cultivation space will be created with around 102,000 sqft for research and development.
Managing director of Peel NRE, part of Peel L&P, Myles Kitcher, said: “This is a game-changing opportunity for the Isle of Man to get ahead in a new and exciting industry that will bring many benefits to the Island and its people and we also hope that it will encourage more renewable projects in the area.
“As expected, our proposals have attracted a lot of attention and, we are pleased to say, significant support from the community during the extensive public consultation. This includes positivity around the emerging industry on the Island, the proposed campus-style development and new educational and career opportunities.
“We will continue to work with the community over the coming weeks and months as the project progresses and will be holding an industry event later in the year for businesses interested in finding out more about available opportunities.”
The proposals (22/00678/B) follow a change in the legislation of cannabis production and exportation on the island. The Isle of Man Government announced in June last year that opening the island up could make it “a world-leading exporter” of cannabis, and establish it as a global destination for science and technological excellence that will contribute to cutting-edge research into cannabinoids for pharmaceutical uses.
It will also set a gold standard for the burgeoning cannabis industry across the world and unlock partnerships in the public and private sectors.
Additionally, the scheme will feature a solar farm to power the site which would be the Island’s only grid-scale renewable project, contributing to the Isle of Man Government’s ambitions on climate change.
Peel NRE launched a public consultation to help shape the plans before the application was submitted to the Department of Environment, Food and Agriculture later this year, and has stated that it has had positive feedback from the Isle of Man community.
Isle of Man Minister for Treasury, Dr Alex Allinson MHK, said: “The development of the Island’s medicinal cannabis sector is a key part of the Government’s ambitions to diversify the economy as well as creating a vibrant new sector to attract more investment and skills to the Isle of Man.
“We are looking to create a world-class infrastructure and continue to welcome interest in developing this sector further.”
Speaking to Cannabis Wealth at the time of the proposal’s announcement, Laurence Skelly, President of Tynwald and Minister for Enterprise, commented: “Diversifying the economy is a significant part of the Isle of Man Government’s Island Plan and the development of Medicinal Cannabis for export is one of the key sectors to bring forward this diversification.
“The Island’s Medicinal Cannabis export proposition is to develop high-quality products and attract new investment through utilising the Island’s stellar reputation as a well-regulated jurisdiction.”
MoU to establish medical cannabis production facility
The site will be based in Madrid, Spain.
Kanabo has signed a Memorandum of Understanding (MoU) for an indoor medical cannabis cultivation project in Spain.
The facility will enable 4,000kg per annum indoor cultivation and processing of cannabis. The facility, which is to be established in Madrid, Spain, will focus exclusively on medical cannabis.
Kanabo Group has formed an Israeli subsidiary company, Kanabo Agritec Ltd. (Agritec), which Kanabo Research Ltd, a wholly owned subsidiary of Kanabo Group, holds a 40 per cent shareholding, together with certain additional control rights over the strategic direction of the subsidiary.
Kanabo Agritec will enter into agreements with customers to offer consulting advice and support on cultivation, processing and production of medical cannabis products to organisations and entrepreneurs entering or already active in the cannabis market.
CEO of Kanabo Group, Avihu Tamir, commented: “We are truly excited by today’s announcement. Agritec provides Kanabo Group plc an opportunity to realise the value of our extensive, existing intellectual capital in medical Cannabis cultivation and production, providing near-term consulting revenues.
“Secondly, it provides us a way to diversify, secure and quality control our supply of medical-grade Cannabis that will always meet our exacting standards, avoiding any wasted margin from product that does not meet the grade.
“In short, the new venture provides all the benefits and value of owning the cultivation supply chain, without the Capex and Opex requirement of building a cultivation operation ourselves. Furthermore, Kanabo consumers will be guaranteed a consistent, highest quality supply of medical Cannabis products, at all times.”
CEO of Agritec, Ophir Shimshi, commented: “We are excited to combine Agritec’s team experience with Kanabo’s extensive R&D knowledge and expertise, to bring this powerful, full-service consultancy offering to market.
“Agritec’s clients will benefit from Kanabo’s immensely valuable, tried, tested and proven playbook.
“Our range of consulting services deliver every strategic and operational consideration required by enterprises and entrepreneurs who wish to take advantage of the immense growth opportunities this market has to offer.”
Kanabo has stated that Agritec provides the company with complementary near-term revenue opportunities and will offer Kanabo improved security of cannabis supply through a diverse range of suppliers, who will all adopt Kanabo’s high-quality manufacturing standards without Kanabo having to fund or become directly involved in cannabis cultivation.
Agritec customers will benefit from Kanabo’s services including: procurement, design and build of commercial scale, medical cannabis cultivation facilities; high-quality genetics and standard operating procedures for optimal medical cannabis cultivation; post-harvest services including offtake agreements with Kanabo Group; design and draft reporting and documentation filing services with the local authorities; and, local training and ongoing consulting services.
As per the terms of the MoU, Agritec may elect to receive up to 20 per cent ownership of the project based on the achievement of agreed milestones.