The present technology provides trichome specific promoters of cannabinoid biosynthesis enzyme genes from Cannabis, nucleotide sequences of the trichome specific promoters, and uses of the promoters for modulating the production of cannabinoids and other compounds in organisms. The present technology also provides chimeric genes, vectors, and transgenic cells and organisms, including plant cells and plants, comprising the trichome specific promoters. Also provided are methods for expressing nucleic acid sequences in cells and organisms using the trichome specific promoters.

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 technology provides trichome specific promoters of cannabinoid biosynthesis enzyme genes from Cannabis, nucleotide sequences of the trichome specific promoters, and uses of the promoters for modulating the production of cannabinoids and other compounds in organisms. The present technology also provides chimeric genes, vectors, and transgenic cells and organisms, including plant cells and plants, comprising the trichome specific promoters. Also provided are methods for expressing nucleic acid sequences in cells and organisms using the trichome specific promoters.

Provided herein is a recombinant vector that comprises a promoter, a secretion tag gene of interest (GOI), and optionally a purification and/or a detection tag. The GOI comprises a polynucleotide sequence of SEQ ID NOs: 2-45 or a polynucleotide sequence with at least 90%, 95%, 98%, 99% or 99.9% identity with any one of SEQ ID NOs: 2-45. Also, disclosed are the sequences of polypeptides that result from expression of a GOI by a host expression system, and a method of producing cannabinoid compounds.

Compositions and methods for providing desired cannabinoid content in cannabis plants. More particularly, the invention relates to compositions and methods for using cannabinoid synthase paralogs as guidance for breeding cannabis plants with a desired cannabinoid content, including but not limited to cultivars, varieties, lines and methods of breeding the same for commercial use.

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.

This document relates to using bidirectional, multi-enzymatic scaffolds to biosynthesize cannabinoids in recombinant hosts.

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 invention provides an apparatus and methods for producing tetrahydrocannabinolic acid (THCA), cannabichromenic acid (CBCA) and cannabichromenic acid (CBCA) in different ratios. The apparatus comprises: (i) a bioreactor comprising (a) an automated supply system configured to deliver a first automated supply of cannabigerolic acid (CBGA), a cannabinoid acid synthase, and a reaction mixture; and (b) a second automated system to cease the reaction; (ii) a controller configured to modify a property of the reaction mixture to produce the desired products; and (iii) an extractor configured to recover the tetrahydrocannabinolic acid (THCA), cannabichromenic acid (CBCA) or cannabidiolic acid (CBDA) and cannabichromenic acid.