Research Projects

Cannabis sativa is presently the forefront of medicinal research due to the significance of cannabinoids. The first plant-derived, pure cannabidiol drug Epidiolex was approved by FDA in 2018. It has been established that two major pathways; polyketide and terpenoid biosynthesis provide substrate flux for cannabinoid biosynthesis. The polyketide backbone of the cannabinoids is derived from olivetolic acid. The biosynthesis of olivetolic acid involves condensation of hexanoyl-CoA with three molecules of malonyl-CoA (Xie et al. 2023). Two sequentially acting enzymes, namely tetrakedide synthase (TKS) and olivetolic acid cyclise (OAC) involved in the biosynthesis of olivetolic acid have been identified and characterized (Gagne et al. 2012). The prenylation of olivetolic acid with geranyldiphosphate leads to the biosynthesis of cannabigerolic acid, catalyzed by an aromatic prenyltransferase. The conversion of cannabigerolic acid to cannabinoids is carried out by enzymes belonging to Berberine Bridge family (Sirikantaramas et al. 2004; Taura et al. 2007). Δ¹-Tetrahydrocannabinolic acid synthase (THCAS) catalyzes the oxidative cyclization of cannabigerolic acid into THCA, which is non-enymzatically de-carboxylated into THC. Similarly, the cyclization of cannabigerolic acid into CBDA is catalyzed by cannabidiolic-acid synthase (CBDAS). The identification of structural genes of cannabinoid biosynthesis provided opportunities to develop alternate production systems for cannabinoids. CSIR-Indian Institute of Integrative Medicine (IIIM), Jammu leads in cannabis-based pharmaceutical research and have received cultivation licenses for R&D. Growing C. sativa is tedious, therefore, it is imperative to develop alternate production technique such as heterologous plant expression, in vitro system, and genome editing. Callus culture is one of the techniques applied in various plants but cannabinoid production (0.0010% - 0.01%) in callus culture of Cannabis is not enough. This study intends to study the molecular aspect of cannabinoid biosynthesis in trichomes and callus tissues and develop genetically modified callus lines for industrial synthesis of cannabinoid. References Gagne, Steve J., et al. "Identification of olivetolic acid cyclase from Cannabis sativa reveals a unique catalytic route to plant polyketides." Proceedings of the National Academy of Sciences 109.31 (2012): 12811-12816. Sirikantaramas, Supaart, et al. "The gene controlling marijuana psychoactivity: molecular cloning and heterologous expression of Δ1-tetrahydrocannabinolic acid synthase from Cannabis sativa L." Journal of Biological Chemistry 279.38 (2004): 39767-39774. Taura, Futoshi, et al. "Cannabidiolic-acid synthase, the chemotype-determining enzyme in the fiber-type Cannabis sativa." FEBS letters 581.16 (2007): 2929-2934. Xie, Ziyan, et al. "Cannabis sativa: origin and history, glandular trichome development, and cannabinoid biosynthesis." Horticulture Research 10.9 (2023): uhad150.

Cannabis sativa, cannabinoid biosynthesis, callus culture, comparative transcriptomics, genetic engineering, secondary metabolites