Embodiments of the disclosure encompass systems, methods, and compositions related to selective advantages to somatic cells that harbor one or more particular genetic modifications. In particular embodiments, there is selective expansion of gene-targeted cells wherein the strategy involves deletion of an essential gene product that is replaced with targeted integration that also includes integration of a therapeutic transgene. The cells that harbor the replaced essential gene product, and thereby the therapeutic transgene, are selected for using pharmaceutical or nutritional agents that are linked to the function of the essential gene product.

The present disclosure provides genetically modified host cells that produce a cannabinoid, a cannabinoid derivative, a cannabinoid precursor, or a cannabinoid precursor derivative. The present disclosure provides methods of synthesizing a cannabinoid, a cannabinoid derivative, a cannabinoid precursor, or a cannabinoid precursor derivative.

The present invention provides for a genetically modified host cell capable of producing isopentenol and/or 3-methyl-3-butenol, comprising (a) an increased expression of phosphomevalonate decarboxylase (PMD) (b) an increased expression of a phosphatase capable of converting isopentenol into 3-methyl-3-butenol, (c) optionally the genetically modified host cell does not express, or has a decreased expression of one or more of NudB, phosphomevalonate kinase (PMK), and/or PMD, and (d) optionally one or more further enzymes capable of converting isopentenol and/or 3-methyl-3-butenol into a third compound, such as isoprene.

Disclosed is a gene multiple insertion cassette set including rDNA NTS fragments and an auxotrophic selection marker having an incomplete promoter is developed, and a safe oral recombinant strain having no antibiotic resistant marker is constructed by multiple insertion of an optimum number of the developed gene multiple insertion cassette sets into chromosomes of a Saccharomyces cerevisiae strain, a vaccine composition including, as an active ingredient, the above strain, a culture product thereof, a cell lysate, or nodavirus capsid protein (NNVcp) isolated and purified therefrom, and a composition for feed addition including, as an active ingredient, the above strain, a culture product thereof, a cell lysate, or squalene or oxidosqualene isolated and purified therefrom.

The present disclosure provides methods of producing commodity products, the methods involving culturing a host cell that is genetically modified to produce a uronate dehydrogenase (UDH) that converts a sugar acid to its corresponding 1,5-aldonolactone, that uses NADP.sup.+ or NAD.sup.+ as a cofactor, and that produces NADPH or NADH, respectively, where the host cell coexpresses an endogenous or a heterologous reductase that utilizes the produced NADPH or NADH to generate the commodity product or a precursor thereof. The present disclosure provides a method of producing downstream products of glycerol and pyruvate in a genetically modified microbial host cell, the method involving culturing a genetically modified microbial host cell of the present disclosure in a culture medium comprising D-galacturonic acid. The present disclosure provides variant UDH polypeptides that utilize NADP.sup.+, nucleic acids encoding the variant UDH polypeptides; and host cells genetically modified with the nucleic acids.

The invention relates generally to materials and methods for biosynthesising quillaic acid in a host by expressing heterologous nucleotide sequences in the host each of which encodes a polypeptide which in combination have said QA biosynthesis activity. Example polypeptides include (i) a Beta-amyrin synthase; (ii) an enzyme capable of oxidising Beta-amyrin or an oxidised derivative thereof at the C-28 position to a carboxylic acid; (iii) an enzyme capable of oxidising Beta-amyrin or an oxidised derivative thereof at the C-16a position to an alcohol; and (iv) an enzyme capable of oxidising Beta-amyrin or an oxidised derivative thereof at the C-23 position to an aldehyde. Preferred nucleotide sequences are obtained from, or derived from, Q. saponaria.

The invention provides a method for producing a terpene or a precursor thereof by microbial fermentation. Typically, the method involves culturing a recombinant bacterium in the presence of a gaseous substrate whereby the bacterium produces a terpene or a precursor thereof, such as mevalonic acid, isopentenyl pyrophosphate, dimethylallyl pyrophosphate, isoprene, geranyl pyrophosphate, farnesyl pyrophosphate, and/or farnesene. The bacterium may comprise one or more exogenous enzymes, such as enzymes in mevalonate, DXS, or terpene biosynthesis pathways.

The present disclosure provides genetically modified host cells that produce a cannabinoid, a cannabinoid derivative, a cannabinoid precursor, or a cannabinoid precursor derivative. The present disclosure provides methods of synthesizing a cannabinoid, a cannabinoid derivative, a cannabinoid precursor, or a cannabinoid precursor derivative.

The invention provides a method for producing a terpene or a precursor thereof by microbial fermentation. Typically, the method involves culturing a recombinant bacterium in the presence of a gaseous substrate whereby the bacterium produces a terpene or a precursor thereof, such as mevalonic acid, isopentenyl pyrophosphate, dimethylallyl pyrophosphate, isoprene, geranyl pyrophosphate, farnesyl pyrophosphate, and/or farnesene. The bacterium may comprise one or more exogenous enzymes, such as enzymes in mevalonate, DXS, or terpene biosynthesis pathways.

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.