Provided are a composition for editing a Cannabis gene, a kit for editing a Cannabis gene, a kit for inhibiting Cannabis DNA expression and targeting the DNA, and a method of editing Cannabis DNA, each using a gene editing protein; a method of preparing an edited Cannabis gene including a step of forming calli; a gRNA for editing a Cannabis gene; and a use of the gRNA of the present invention and a use of both the gene editing protein and the gRNA for editing a Cannabis gene.

The present disclosure provides methods for making glycosylated cannabinoids including methods using recombinant host cells comprising a pathway capable of producing a cannabinoid and a heterologous nucleic acid that encodes a UDP-glycosyl transferase. The disclosure also provides compositions of recombinant host cells capable of producing glycosylated cannabinoids, and compositions and uses of the glycosylated cannabinoids.

The present disclosure relates generally to methods, isolated polypeptides and polynucleotides, expression vectors, and host cells for the production of cannabidiolic acid (CBDa), cannabigerolic acid (CBCa), and other phytocannabinoids. A method of producing CBDa, CBCa, and/or other phytocannabinoids in a heterologous host cell having CBDa-producing, CBCa-producing or phytocannabinoid-producing capacity comprises transforming the host cell with a nucleotide encoding a variant CBDa synthase protein having a serine insertion between residues P224 and K225 and one or more other amino acid mutation relative to wild type CBDa synthase, and culturing the transformed host cell to produce CBDa, CBCa, and/or other phytocannabinoids therefrom. The variant CBDa synthase protein has at least 85% sequence identity with the wild type CBDa synthase protein sequence OXC52 according to SEQ ID NO:140, with serine insertion (SEQ ID NO:141). Exemplary variants having good phytocannabinoid production capacity are described.

The present disclosure relates to the production of cannabinoids in yeast. In as aspect there is provided a genetically modified yeast comprising: one or more GPP producing genes and optionally, one or more GPP pathway genes; two or more olivetolic acid producing genes; one or more cannabinoid precursor or cannabinoid producing genes; one or more Hexanoyl-CoA producing genes, and at least 5% dry weight of fatty acids or fats.

The present invention provides methods and compositions for producing cannabinoids in photosynthetic microorganisms, e.g., cyanobacteria.

Methods and systems for the alteration of cannabinoid expression and composition are described. Modular systems for processing cannabis and hemp are described. Described components of the modular system include improved recovery of high-value cannabis components and methods for utilizing residual biomass components after the cannabis plant has been fully processed.

Aspects of the disclosure relate to biosynthesis of cannabinoids and cannabinoid precursors in recombinant cells and in vitro.

A method of increasing the cannabinoid levels in a genetically modified Cannabis sativa plant includes genetically modifying the plant to induce the overexpression of the gene that controls the expression of tetrahydrocannabinolic acid (THCA) synthase and/or cannabidiolic acid (CBDA) synthase. The overexpression of THCA synthase and CBDA synthase catalyzes an increased synthesis of cannabigerolic acid to tetrahydrocannabinolic acid and cannabidiolic acid, as well as the cannabinoids (3aR)-2,4,4-trimethyl-7-pentyl-3,3a,4,9b-tetrahydrocyclopenta[c]chromen-9-ol and 2-((1R,5R)-3-methyl-5-(prop-1-en-2-yl)cyclopent-2-en-1-yl)-5-pentylbenzene-1,3-diol. Pharmaceutical compositions comprising the modified cannabinoids produced by the transgenic Cannabis sativa plant or prepared synthetically are used to treat various diseases and conditions.

The present disclosure provides methods and kits for characterizing Cannabis plants. Methods and kits of the present disclosure include detection/amplification of one or more enzymes involved in the production of cannabinoids, such as, for example, tetrahydrocannabinolic acid (THCA) synthase and/or cannabidiolic acid (CBDA) synthase.

The methods and systems disclose enzymes that function to import malonic acid or malonates into a cell. The enzymes increase the output of precursor molecules by enriching certain pathways in the cell. The precursor molecules can be converted to cannabinoids. The enzymes are a family of proteins which have a majority of common alignments.