Provided is a Cannabis plant with reduced delta-9-tetrahydrocannabinol (THC) content, or reduced cannabidiol (CBD) content, or reduced THC and CBD content. According to a core aspect of the invention, the Cannabis plant comprises at least one targeted genome modification effective in decreasing expression of. at least one Cannabis gene encoding a cannabinoid biosynthesis enzyme selected from the group consisting of Cannabis tetrahydrocannabinolic acid synthase (CsTHCAS), Cannabis cannabidiolic acid synthase (CsCBDAS), Cannabis aromatic prenyltransferase (CsPT), Cannabis olivetol synthase (CsOLS), Cannabis acyl-activating enzyme 1 (CsAAE1) and any combination thereof. Further disclose are methods for production of the Cannabis plants and use thereof.

Enzymes involved in cannabinoid biosynthesis are recombinantly expressed in a host cell. The host cell may be a prokaryote (e.g. Escherichia coli) or a eukaryote (e.g. Yarrowia lipolytica). The enzymes include a heterologous cannabigerolic acid synthase as well as additional enzymes involved in the biosynthesis of cannabinoid precursors such as geranyl diphosphate, olivetol, olivetolic acid, divarin and/or divarinic acid. Methods are provided for producing C5-cannabinoids and/or C3-cannabinoids by fermentation of the recombinant host cell. Alternatively, cannabinoids can be produced by biotransformation of cannabinoid precursors in recombinant cells or by disrupted recombinant cells.

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.

A method is provided for biosynthetic production of cannabinoids in microorganisms from a carbon source precursor. This method describes the genetic modifications needed to engineer microorganisms to produce cannabinoids as well as a method for identifying and quantifying cannabinoids from fermentation broth. A system is also provided for tuning the method to produce different cannabinoids of interest by systematically modulating the enzymes encoded by the genetic modifications introduced in the microorganism.

Exemplary embodiments provided herein include genetically engineering microorganisms, such as yeast or bacteria, to produce cannabinoids by inserting genes that produce the appropriate enzymes for the metabolic production of a desired compound.

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.

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.