Abstract: The present disclosure generally relates to novel polynucleotide molecules for use in regulating gene expression in recombinant cells, such as labyrinthulomycetes cells. The disclosure further relates to nucleic acid constructs, such as vectors and expression cassettes, containing a regulatory element operably linked to a heterologous nucleotide sequence. The disclosure further relates to methods for stably transforming a host cell, such as a labyrinthulomycetes cell with transgenes. Stably transformed recombinant cells, progeny, biomaterials derived therefrom, and methods for preparing and using the same are also provided.

Abstract: The invention provides a recombinant Thraustochytriaceae cell for the production of a glycomolecule. The cell comprises a nucleic acid encoding a heterologous glycomolecule, and a sequence encoding a heterologous oligosaccharyltransferase. The cell produces the heterologous glycomolecule having a higher glycan occupancy compared to the same heterologous glycomolecule produced by a corresponding cell not comprising the heterologous oligosaccharyltransferase. The glycan occupancy can be more than 25%. The cells advantageously produce and, optionally secrete, the heterologous glycomolecule. Thus, the invention provides recombinant organisms that provide glycomolecules having a glycosylation profile that is more similar to the glycosylation profile produced in a manunalian cell.

Abstract: The invention provides a recombinant Labyrinthulomycetes cell for the production of a low sulfate glycomolecule. The cell comprises a nucleic acid encoding a heterologous glycomolecule, and a sequence encoding a heterologous oligosaccharyltransferase. The cell produces the heterologous glycomolecule having fewer sulfated glycans compared to the same heterologous glycomolecule produced by a corresponding cell not comprising the heterologous oligosaccharyltransferase. The cells advantageously produce and, optionally secrete, the heterologous glycomolecule. Thus, the invention provides recombinant organisms that provide glycomolecules having a glycosylation profile that is more similar to the glycosylation profile produced in mammalian cell.

Abstract: The invention provides recombinant organisms containing a nucleic acid encoding a heterologous glycomolecule that has a low sulfation profile or that is unsulfated. In one embodiment the heterologous glycomolecule is an immunoglobulin molecule. The recombinant organisms have a genetic modification to at least one sulfotransferase gene, such as a deletion, disruption, or other genetic modification. The cells advantageously produce and, optionally secrete, the heterologous glycomolecule. Thus, the invention provides recombinant organisms that provide glycomolecules having a glycosylation profile that is more similar to the glycosylation profile produced in a mammalian cell, and therefore may be safer and more effective for use as a therapeutic in humans or animals. The glycomolecules can be a glycoprotein, glycopeptide, or a glycolipid.

Abstract: The present disclosure generally relates to novel polynucleotide molecules for use in regulating gene expression in recombinant cells, such as labyrinthulomycetes cells. The disclosure further relates to nucleic acid constructs, such as vectors and expression cassettes, containing a regulatory element operably linked to a heterologous nucleotide sequence. The disclosure further relates to methods for stably transforming a host cell, such as a labyrinthulomycetes cell with transgenes. Stably transformed recombinant cells, progeny, biomaterials derived therefrom, and methods for preparing and using the same are also provided.

Abstract: The present disclosure generally relates to novel polynucleotide molecules for use in regulating gene expression in recombinant cells, such as labyrinthulomycetes cells. The disclosure further relates to nucleic acid constructs, such as vectors and expression cassettes, containing a regulatory element operably linked to a heterologous nucleotide sequence. The disclosure further relates to methods for stably transforming a host cell, such as a labyrinthulomycetes cell with transgenes. Stably transformed recombinant cells, progeny, biomaterials derived therefrom, and methods for preparing and using the same are also provided.

Abstract: The present invention provides recombinant host cells that produce proteins or therapeutic proteins, and nucleic acid constructs for producing the cells. The cells have nucleic acid constructs that encode a heterologous protein, for example an antibody. The nucleic acid constructs also can have a functional signal sequence that directs the secretion of the protein from the cell. The signal sequence can be any functional signal sequence, and various signal sequences are disclosed herein. The invention also provides methods of producing the proteins.

Abstract: The present invention discloses methods for assembling a nucleic acid molecule from a set of overlapping oligonucleotides. The method involves contacting a set of overlapping oligonucleotides with a DNA polymerase, a mixture of dNTPs, and a crowding agent to form an assembly mixture. In one embodiment the crowding agent is polyethylene glycol (PEG). The presence of the crowding agent facilitates the nucleic acid assembly process of the invention. The assembly mixture is then subjected to multiple cycles, each cycle comprising an annealing phase, an extension phase, and a denaturation phase, and the desired nucleic acid molecule is thereby assembled. In some embodiments one or more of the phases are time varied.

Abstract: The present disclosure relates in part to recombinant microorganisms that include non-native genes encoding PUFA-PKS polypeptides, and to methods of making and using such microorganisms for producing at least one PUFA. In particular, the disclosure further relates to methods and related materials useful for the production of at least one PUFA by heterologous expression of the nucleic acid sequences disclosed herein encoding PUFA-PKS polypeptides.

Abstract: The present disclosure generally relates to novel polynucleotide molecules for use in regulating gene expression in recombinant cells, such as labyrinthulomycetes cells. The disclosure further relates to nucleic acid constructs, such as vectors and expression cassettes, containing a regulatory element operably linked to a heterologous nucleotide sequence. The disclosure further relates to methods for stably transforming a host cell, such as a labyrinthulomycetes cell with transgenes. Stably transformed recombinant cells, progeny, biomaterials derived therefrom, and methods for preparing and using the same are also provided.

Abstract: The present invention provides recombinant microorganisms comprising an isobutanol producing metabolic pathway and methods of using said recombinant microorganisms to produce isobutanol. In various aspects of the invention, the recombinant microorganisms may comprise a modification resulting in the reduction of pyruvate decarboxylase and/or glycerol-3-phosphate dehydrogenase activity. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.

Abstract: The present disclosure generally relates to novel polynucleotide molecules for use in regulating gene expression in recombinant cells, such as labyrinthulomycetes cells. The disclosure further relates to nucleic acid constructs, such as vectors and expression cassettes, containing a regulatory element operably linked to a heterologous nucleotide sequence. The disclosure further relates to methods for stably transforming a host cell, such as a labyrinthulomycetes cell with transgenes. Stably transformed recombinant cells, progeny, biomaterials derived therefrom, and methods for preparing and using the same are also provided.

Abstract: The present invention provides recombinant microorganisms comprising an isobutanol producing metabolic pathway and methods of using said recombinant microorganisms to produce isobutanol. In various aspects of the invention, the recombinant microorganisms may comprise a modification resulting in the reduction of pyruvate decarboxylase and/or glycerol-3-phosphate dehydrogenase activity. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.

Abstract: The present invention provides recombinant host cells that produce proteins or therapeutic proteins, and nucleic acid constructs for producing the cells. The cells have nucleic acid constructs that encode a heterologous protein, for example an antibody. The nucleic acid constructs also can have a functional signal sequence that directs the secretion of the protein from the cell. The signal sequence can be any functional signal sequence, and various signal sequences are disclosed herein. The invention also provides methods of producing the proteins.

Abstract: The present application provides novel regulatory elements including promoter sequences from marine microorganisms. The application further discloses DNA constructs containing these novel regulatory elements; transgenic cells, transgenic non-human organisms, and progeny containing these novel regulatory elements. Methods of modifying, producing, and using the regulatory elements are also disclosed. The regulatory elements disclosed herein are particularly suited for use in Nannochloropsis and other microalgae.

Abstract: The present invention relates to recombinant microorganisms comprising biosynthetic pathways and methods of using said recombinant microorganisms to produce various beneficial metabolites. In various aspects of the invention, the recombinant microorganisms may further comprise one or more modifications resulting in the reduction or elimination of 3 keto-acid (e.g., acetolactate and 2-aceto-2-hydroxybutyrate) and/or aldehyde-derived by-products. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces Glade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.

Abstract: Recombinant microorganisms engineered for the production of polyunsaturated fatty acids (PUFAs) are provided. Also provided are biomass, microbial oils, and food products and ingredients produced by or comprising the microorganisms of the invention.

Abstract: Methods and materials useful for modulating the sensitivity of cells to an inhibitor of acetohydroxyacid synthase (AHAS) are disclosed. For example, nucleic acid molecules encoding AHAS large subunits are disclosed as well as methods for using such nucleic acid molecules to transform microbial cells and plant cells, and to confer modulated sensitivity to AHAS-inhibiting compounds onto such cells. Further provided are materials and methods useful for modulating growth, development, activity, and characteristics of host cells and organisms.

Abstract: The present disclosure generally relates to novel polynucleotide molecules for use in regulating gene expression in recombinant cells, such as labyrinthulomycetes cells. The disclosure further relates to nucleic acid constructs, such as vectors and expression cassettes, containing a regulatory element operably linked to a heterologous nucleotide sequence. The disclosure further relates to methods for stably transforming a host cell, such as a labyrinthulomycetes cell with transgenes. Stably transformed recombinant cells, progeny, biomaterials derived therefrom, and methods for preparing and using the same are also provided.

Abstract: The present disclosure relates in part to recombinant microorganisms that include non-native genes encoding PUFA-PKS polypeptides, and to methods of making and using such microorganisms for producing at least one PUFA. In particular, the disclosure further relates to methods and related materials useful for the production of at least one PUFA by heterologous expression of the nucleic acid sequences disclosed herein encoding PUFA-PKS polypeptides.