Synthetic Genomics, Inc.

Viridos, Inc.

Exxonmobil Research and Engineering Company

John Verruto

Eric R Moellering

Imad Ajjawi

Moena Aqui

Erik Holtzapple

Roberto Spreafico

Randor R Radakovits

Fedor I Kuzminov

Andrew Schultz

Saheed Imam

Nicholas Bauman

Kathleen Kwok

Shaun Bailey

Jennifer Nominati

Stephen K Picataggio

Paul Gordon Roessler

Robert Christopher Brown

Bo Liu

Kevin T Watts

Jane C Schneider

Rekha Seshadri

Kevin V Martin

Soyan Leung Lieberman

William F Lambert

Jennifer Coppersmith

Sarah Potts

Jessica N Weir

Jessica Greiner

Amanda Skaggs

Jessica Weir

Robert C Brown

The invention provides methods of transforming photosynthetic organisms, such as green algae. The methods involve methylating one or more DNA fragments of a DNA construct and transforming the one or more fragments into the photosynthetic organism. The DNA fragments can be the product of a DNA amplification procedure, such as PCR or a PCR-like procedure. In one embodiment the one or more fragments of DNA that comprise a DNA construct are dam methylated prior to being transformed into the photosynthetic organism.

Method of transforming photosynthetic organisms

The present invention provides cell lines for high efficiency genome editing using cas/CRISPR systems, methods of generating such cells lines and methods of generating mutations in the genome of an organism using such cell lines.

RNA-guided endonuclease expressing algal strain for high efficiency in vivo genome editing

Disclosed herein are mutant photosynthetic microorganisms having an attenuated SGI1 gene. The mutants have reduced chlorophyll and increased productivity with respect to wild type cells. Also disclosed are methods of using such mutants for producing biomass or bioproducts, and methods of screening for such mutants.

High productivity algal mutants having reduced photosynthetic antenna

The invention provides important tools for the use of Chlorophyte organisms, including regulatory sequences useful for the expression of transgenes in such organisms. Also provided are vectors and expression cassettes containing promoters and/or terminators disclosed herein for expression of transgenes in Chlorophyte organisms. Methods of using these tools are also provided.

Regulatory sequences for expression of transgenes

The present invention provides cell lines for high efficiency genome editing using cas/CRISPR systems, methods of generating such cells lines, and methods of generating mutations in the genome of an organism using such cell lines.

Compositions and methods for high efficiency in vivo genome editing

Mutant photosynthetic organisms having reduced chlorophyll and increased photosynthetic efficiency are provided. The mutant strains have mutated or attenuated: chloroplastic SRP54 gene and SGI1 gene; chloroplastic SRP54 gene and SGI2 gene; chloroplastic SRP54 gene, SGI1, and SGI2 genes are disclosed. The mutant photosynthetic organisms exhibit increased productivity with respect to wild-type strains. Also provided are mutant photosynthetic organisms having mutated or attenuated cytosolic SRP54 genes. Provided herein are methods of producing biomass and other products such as lipids using strains having mutations in an SRP54 gene, SGI1, SGI2 genes, a combination of SGI1/SRP54, and a combination of SGI2 and SRP54 genes. Also included are constructs and methods for attenuating or disrupting SRP54, SGI1, and SGI2 genes.

Genetic modulation of photosynthetic organisms for improved growth

The present application provides novel algal regulatory elements including inducible nitrate/nitrite promoter sequences and terminator sequences. The application further discloses DNA constructs comprising these novel regulatory elements, and recombinant microorganisms comprising these regulatory elements. Methods of modifying, producing, and using the regulatory elements are also disclosed. Methods disclosed in the present application are suited for inducible expressions of genes, such as a transgene or a native gene in algal species.

Inducible expression of genes in algae

The present invention provides mutant microorganisms having attenuated expression of a gene encoding a polypeptide that includes a TPR domain, wherein the mutant microorganisms have higher lipid productivity and/or exhibit increased partitioning of carbon to lipid as compared to wild type microorganisms from which they are derived. Also provided are methods of producing lipids using the mutant microorganisms, guide RNAs, and nucleic acid constructs used for producing mutant microorganisms.

Algal lipid productivity via genetic modification of a TPR domain containing protein

Disclosed herein are mutant photosynthetic microorgnaisms having an attenuated SGI1 gene. The mutants have reduced chlorophyll and increased productivity with respect to wild type cells. Also disclosed are methods of using such mutants for producing biomass or bioproducts, and methods of screening for such mutants.

The invention relates to acyl-CoA-independent methods of producing fatty alcohols in recombinant host cells engineered to express an alcohol-forming acyl-ACP reductase. The recombinant host cells may be photosynthetic microorganisms, such as cyanobacteria. Isolated nucleic acid molecules, vectors, and recombinant host cells expressing an alcohol-forming acyl-ACP reductase, and systems for producing fatty alcohols via an acyl-CoA-independent pathway, are also provided. Also provided are microorganisms engineered for the secretion of fatty acids and fatty acid derivatives, including fatty alcohols, and methods of producing fatty acid derivatives using such engineering microorganisms.

Production and secretion of fatty acids and fatty acid derivatives

The invention relates to methods for producing a wax ester in recombinant host cells engineered to express a thioesterase, an acyl-CoA synthetase, an alcohol-forming fatty acyl reductase, and a wax ester synthase. The methods of the invention may take place in photosynthetic microorganisms, and particularly in cyanobacteria. Isolated nucleotide molecules and vectors expressing the thioesterase, acyl-CoA synthetase, alcohol-forming fatty acyl reductase, and wax ester synthase, recombinant host cells expressing the thioesterase, acyl-CoA synthetase, alcohol-forming fatty acyl reductase, and wax ester synthase, and systems for producing a wax ester via a pathway using these four enzymes, are also provided.

Four-gene pathway for wax ester synthesis

The invention relates to acyl-CoA-independent methods of producing a wax ester in recombinant host cells engineered to express an acyl-ACP wax ester synthase, and an alcohol-forming acyl-ACP reductase. The methods of the invention may take place in photosynthetic microorganisms, and particularly in cyanobacteria. Isolated nucleotide molecules and vectors expressing an acyl-ACP wax ester synthase and/or an alcohol-forming acyl-ACP reductase, recombinant host cells expressing an acyl-ACP wax ester synthase and optionally an alcohol-forming acyl-ACP reductase, and systems for producing a wax ester via an acyl-CoA-independent pathway, are also provided.

Acyl-ACP wax ester synthases

The present invention provides novel promoter and terminator sequences for use in gene expression in eukaryotic cells, such as algal cells. The invention further provides expression cassettes comprising a promoter, as described herein, operably linked to a gene. The invention further provides expression vectors and host eukaryotic cells, such as algal cells, for expressing a protein encoded by the gene; and methods for stably transforming eukaryotic algae such aswith transgenes.

Tetraselmis promoters and terminators for use in eukaryotic cells

This invention describes genes, metabolic pathways, microbial strains and methods to produce 2,6-dimethyloctane as an advanced biofuel from renewable feedstocks.

Growing community of inventors

San Diego, CA, United States of America

John Verruto