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.

In an aspect, the disclosure provides methods for making neurotransmitters in a host organism. The neurotransmitters can be cannabinoids and derivatives of cannabinoids. The host cells can be microalgae, fungi or other host cells. In a related aspect, the disclosure provides host cells engineered to have biochemical pathways for making neurotransmitters such as cannabinoids.

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

The invention provides engineered biosynthetic pathways that can be used to produce cannabinoids from fatty acids, recombinant microorganisms incorporating such pathways, methods of biosynthetically producing cannabinoids from fatty acids, and cannabinoids so produced.

The present disclosure provides recombinant host cells comprising a pathway capable of producing a cannabinoid and/or cannabinoid precursor, wherein the pathway comprises an enzyme AAE from a source organism other than Cannabis sativa, such as Humulus lupulus. The disclosure also provides methods of using the host cells to produce rare cannabinoids and/or rare cannabinoid precursors.

Disclosed herein are microorganism and methods that can be used for the synthesis of cannabigerolic acid (CBGA) and cannabinoids. The methods disclosed can be used to produce CBGA, Δ.sup.9-tetrahydrocannabinolic acid (THCA), cannabidiolic acid (CBDA), cannabichromenic acid (CBGA), Δ.sup.9-tetrahydrocannabivarinic acid (THCVA), cannabidivarinic acid (CBDVA), cannabichromevarinic acid (CBCVA), Δ.sup.9-tetrahydrocannabinol (THC), cannabidiol (CBD), cannabichromene (CBC). Enzymes useful for the synthesis of CBGA and cannabinoids, include but are not limited to acyl activating enzyme (AAE1), polyketide synthase (PKS), olivetolic acid cyclase (OAC), prenyltransferase (PT), THCA synthase (THCAS), CBDA synthase (CBDAS), CBC A synthase (CBCAS), HMG-Co reductase (HMG1), and/or famesyl pyrophosphate synthetase (ERG20). The microorganisms can also have one or more genes disrupted, such as gene that that controls beta oxidation of long chain fatty acids.

A nucleic acid molecule is disclosed for transiently transforming a plant to produce Δ9-tetrahydrocannabinolic acid synthase, cannabidiolic acid synthase, and/or cannabichromenic acid synthase. The nucleic acid molecule corresponds to a nucleotide sequence comprising at least one of a nucleotide sequence fragment encoding a polypeptide having at least 78% sequence identity to SEQ ID NO: 4, SEQ ID NO:5 or SEQ ID NO:6 or comprising at least 15 contiguous nucleotides of the nucleotide sequence SEQ ID NO: 4, SEQ ID NO:5 or SEQ ID NO:6. The nucleotide sequence further comprises a KDEL or HDEL retrieval tag for targeting the nucleotide sequence to the endoplasmic reticulum. Further aspects relate to a viral vector comprising such nucleic acid molecule, and to methods for producing THCAS, CBDAS, CBCAS, THCA, CBDA, CBCA, THC, CBD and/or CBC based on transient expression of the nucleic acid sequence in a host plant.

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

Provided is a Cannabis plant exhibiting altered tetrahydrocannabinolic acid (THCA) and/or cannabidiolic acid (CBDA) content. The plant includes a modified genomic locus involved in tetrahydrocannabinolic acid synthase (THCAS) and/or cannabidiolic acid synthase (CBDAS) gene expression, the genomic locus includes at least one targeted nucleotide modification within a regulatory region modulating the expression of at least one allele of the THCAS and/or CBDAS genes. Further provided are methods for producing the aforementioned Cannabis plant.

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.