Abstract: The present invention provides compositions and methods for the production and delivery of recombinant double-stranded RNA molecules (dsRNA) encoding heterologous proteins, which can be useful for various therapeutic purposes as well as for the production of desired proteins. The compositions contain engineered double-stranded RNA particles (dsRPs) that can contain a double-stranded RNA molecule that can be a genome or portion of a genome, which can be enclosed in a capsid or coat protein. The dsRNA molecule also comprises an RNA sub-sequence that encodes a heterologous protein. The dsRPs can be derived from wild-type viral organisms. The delivery of the dsRPs (or DNA or RNA molecules) of the invention to an organism provides for therapeutic benefits as well as for the production of desired proteins.

Abstract: New influenza donor strains for the production of reassortant influenza B viruses are provided.

Abstract: Recombinant photosynthetic microorganisms that convert inorganic carbon to secreted fatty acids are described. Methods to recover the secreted fatty acids from the culture medium without the need for cell harvesting are also described.

Abstract: The present invention provides mutant microorganism that have higher lipid productivity than the wild type microorganisms from which they are derived while biomass at levels that are within approximately 50% of wild type biomass productivities under nitrogen replete conditions. Particular mutants produce at least twice as much FAME lipid as wild type while producing at least 75% of the biomass produced by wild type cells under nitrogen replete conditions. Also provided are methods of producing lipid using the mutant strains.

Abstract: The present invention provides compositions and methods for the production and delivery of recombinant double-stranded RNA molecules (dsRNA) targeting pathogen sequences, which can be useful as an RNA vaccine. The compositions contain engineered double-stranded RNA particles (dsRPs) that can contain a double-stranded RNA molecule that can be a genome or portion of a genome, which can be enclosed in a capsid or coat protein. The dsRNA molecule also comprises an RNA sub-sequence that binds to a target sequence of a pathogenic organism. The dsRPs can be derived from wild-type viral organisms. The delivery of the dsRPs of the invention to an organism provides a protection to the organism from the pathogen.

Abstract: Mutant photosynthetic algae having increased biomass productivity are provided. The mutants have attenuated expression of violaxanthin chlorophyll a binding proteins (VCP) or fucoxanthin chlorophyll a/c binding proteins (FCP), reduced chlorophyll, higher apparent ETR(II), little to no reduction in Pmax per cell, and decreased NPQ over a wide range of light intensities. Provided herein are constructs for attenuating or disrupting VCP or FCP genes. Also provided are methods of culturing VCP or FCP mutants for the production of biomass or other products.

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 invention provides methods of method of synthesizing 2,5-furan dicarboxylic acid (FDCA) and FDCA precursor molecules. The methods involve performing a chemical dehydration reaction on a gluconic acid derivative in the presence of a dehydration catalyst. In some embodiments the gluconic acid derivative can be 2-dehydro-3-deoxy gluconic acid (DHG) or an ester thereof, 2-ketogluconic acid (2KGA) or an ester thereof, and 5-ketogluconic acid (5KGA) or an ester thereof. The 2,5-furan dicarboxylic acid precursor molecule is thereby synthesized, which can be converted into FDCA. The chemical dehydration can be performed by a variety of acid basic catalysts.

Abstract: The present application provides novel regulatory elements including promoter sequences from microorganisms. The application further discloses DNA constructs containing these novel regulatory elements, and recombinant microorganisms comprising these regulatory elements. Methods of modifying, producing, and using the regulatory elements are also disclosed. The regulatory elements and transformation methods disclosed herein are particularly suited for use in Parachlorella and other microalgae.

Abstract: Methods are provided for constructing a synthetic genome, comprising generating and assembling nucleic acid cassettes comprising portions of the genome, wherein at least one of the nucleic acid cassettes is constructed from nucleic acid components that have been chemically synthesized, or from copies of the chemically synthesized nucleic acid components. In one embodiment, the entire synthetic genome is constructed from nucleic acid components that have been chemically synthesized, or from copies of the chemically synthesized nucleic acid components. Rational methods may be used to design the synthetic genome (e.g., to establish a minimal genome and/or to optimize the function of genes within a genome, such as by mutating or rearranging the order of the genes). Synthetic genomes of the invention may be introduced into vesicles (e.g., bacterial cells from which part or all of the resident genome has been removed, or synthetic vesicles) to generate synthetic cells.

Abstract: The invention provides a tamper resistant assembly that that securely contains a valuable material. The assembly has a container for holding the valuable material, an optional carrier that contains the container, one or more cover components that enclose the valuable material in the container, and one or more labels having a plurality of devices that reveal tampering by distortion of at least one of the plurality of devices. The label(s) are positioned so that dislodging a cover component causes a detectable distortion in at least one of the plurality of devices, thereby revealing tampering with the assembly. In one embodiment the label can be affixed partially to a surface of a cover component and partially to a surface of the container or the optional carrier. Also disclosed are methods of detecting tampering and method of manufacturing a temper-resistant assembly.

Abstract: Mutant photosynthetic microorganisms having reduced chlorophyll and increased photosynthetic efficiency are provided. The mutants have a locked in high light-acclimated phenotype, in which many of the photosynthetic parameters characteristic of high light acclimated wild type cells are found in the LIHLA mutants when acclimated to low light, such as reduced chlorophyll, reduced NPQ, higher qP, higher Ek, higher Pmax per unit chlorophyll with little to no reduction in Pmax per cell, and higher rates of electron transport through photosystem II over a wide range of light intensities. Provided herein are constructs for attenuating or disrupting genes are provided for generating mutants having the LIHLA phenotype. Also provided are methods of culturing LIHLA mutants for the production of biomass or other products.

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 novel genes encoding Class II acyl-ACP thioesterases and variants thereof that are active on C8, C10, C12, C14, C16, and C18 acyl-ACP substrates. The thioesterases can be introduced into transgenic organisms, including microorganisms and photosynthetic organisms, for producing fatty acids and fatty acid products.

Abstract: The present invention relates to the culture and manipulation of microorganisms for biotech applications, and is based on the discovery and characterization of spliced leader sequences identified in transcripts from Nannochloropsis species. In particular, the invention provides nucleic acid compositions comprising a SL sequence operably linked to a protein-encoding gene. Further provided are compositions and methods for enhanced gene expression in recombinant microorganisms as well as methods for identification and/or isolation of nucleic acid molecules tagged with a spliced leader sequence.

Abstract: The present invention provides methods of producing dicarboxylic acids. The methods involve incubating a fatty acid or hydrocarbon substrate with an enzyme to produce a dicarboxylic acid product. The enzyme acts on the substrate to produce a product that has been both over-oxidized and has undergone cleavage of a C—C bond. In some embodiments the enzymes having these useful characteristics are mutants of a cytochrome P450 enzyme, for example an enzyme of the class CYP102 or a mutant thereof. The invention provides enzymes where these desirable characteristics can be found in a single enzyme, and thus in some embodiments the methods can be performed through the action of a single enzyme.

Abstract: The present invention provides materials and methods useful for error correction of nucleic acid molecules. In one embodiment of the invention, a first plurality of double-stranded nucleic acid molecules having a nucleotide mismatch are fragmented by exposure to a molecule having unidirectional mismatch endonuclease activity. The nucleic acid molecules are cut at the mismatch site or near the mismatch site, leaving a double-stranded nucleic acid molecule having a mismatch at the end or near end of the molecule. The nucleic acid molecule is then exposed to a molecule having unidirectional exonuclease activity to remove the mismatched nucleotide. The missing nucleotides can then be filled in by the action of, e.g., a molecule having DNA polymerase activity. The result is double-stranded nucleic acid molecules with a decreased frequency of nucleotide mismatches.

Abstract: The disclosure generally relates to methods and materials for modulating cell productivity. In particular, the present disclosure provides polynucleotides encoding transcription factor proteins that when overexpressed in microorganisms result in increased in productivity, such as increased biomass productivity. Also disclosed are methods of using the genetically engineered host strains to modulate or increase productivity of host cells such as, for example, algal or heterokont cells. Genetically engineered host cells, such as algal and heterokont cells having increased biomass productivity and bioproducts derived from such host cells are also disclosed.

Abstract: Provided herein are methods for identifying centromeres and centromeres identified by such methods. Centromeres of organisms such as algae, fungi, and protists can be used, for example, for constructing artificial chromosomes and cells containing such artificial chromosomes.

Abstract: The invention provides a tamper resistant assembly that that securely contains a valuable material. The assembly has a container for holding the valuable material, an optional carrier that contains the container, one or more cover components that enclose the valuable material in the container, and one or more labels having a plurality of devices that reveal tampering by distortion of at least one of the plurality of devices. The label(s) are positioned so that dislodging a cover component causes a detectable distortion in at least one of the plurality of devices, thereby revealing tampering with the assembly. In one embodiment the label can be affixed partially to a surface of a cover component and partially to a surface of the container or the optional carrier. Also disclosed are methods of detecting tampering and method of manufacturing a temper-resistant assembly.