The present disclosure relates to methods, compositions and kits for synthesizing moderate length RNAs (mlRNAs, including gRNAs) by splint-mediated ligation of RNA fragments. The synthesis of moderate length RNAs can be followed by DNase treatment. In some embodiments, splint DNA oligonucleotides that are no longer than 32 nucleotides are used.

The present disclosure relates to an adenylosuccinate synthetase variant, a microorganism containing the same, and a method for preparing purine nucleotides using the microorganism.

Disclosed are compositions and methods for gene editing. The present disclosure relates to compositions and methods for gene editing using a Cas nickase to cleave a double-stranded nucleic acid sequence near a target site and a ligase to incorporate a nucleic acid into a double-stranded nucleic acid sequence. The present disclosure also provides reagents for use in the gene editing methods. The present disclosure further provides kits containing reagents for use in the gene editing methods.

The use of microorganisms to make alpha-functionalized chemicals and fuels, (e.g. alpha-functionalized carboxylic acids, alcohols, hydrocarbons, amines, and their beta-, and omega-functionalized derivatives), by utilizing an iterative carbon chain elongation pathway that uses functionalized extender units. The core enzymes in the pathway include thiolase, dehydrogenase, dehydratase and reductase. Native or engineered thiolases catalyze the condensation of either unsubstituted or functionalized acyl-CoA primers with an alpha-functionalized acetyl-CoA as the extender unit to generate alpha-functionalized β-keto acyl-CoA. Dehydrogenase converts alpha-functionalized β-keto acyl-CoA to alpha-functionalized β-hydroxy acyl-CoA. Dehydratase converts alpha-functionalized β-hydroxy acyl-CoA to alpha-functionalized enoyl-CoA. Reductase converts alpha-functionalized enoyl-CoA to alpha-functionalized acyl-CoA. The platform can be operated in an iterative manner (i.e. multiple turns) by using the resulting alpha-functionalized acyl-CoA as primer and the aforementioned alpha-functionalized extender unit in subsequent turns of the cycle. Termination pathways acting on any of the four alpha-functionalized CoA thioester intermediates terminate the platform and generate various alpha-functionalized carboxylic acids, alcohols and amines with different β-reduction degree.

The invention includes a mutant T4 DNA ligase or a biologically active fragment thereof, which has greater activity than wild type T4 DNA ligase. The mutant T4 DNA ligase, or the biologically active fragment, has one or more substitutions differing from the wild type, as described more fully in the Summary with reference to the Sequence Listing.

Embodiments of the present disclosure pertain to modified plants or seeds that include: (1) overexpressed rice SUMO E3 ligase SIZ1 (OsSIZ1), an analog thereof, a homolog thereof, a derivative thereof, or combinations thereof; and (2) overexpressed Larrea tridentate rubisco activase (LtRCA), an analog thereof, a homolog thereof, a derivative thereof, or combinations thereof. The modified plant or seed demonstrates enhanced resistance to environmental stress. Additional embodiments of the present disclosure pertain to methods of developing a modified plant or seed of the present disclosure by overexpressing in the plant or seed: (1) rice SUMO E3 ligase SIZ1 (OsSIZ1), an analog thereof, a homolog thereof, a derivative thereof, or combinations thereof; and (2) Larrea tridentate rubisco activase (LtRCA), an analog thereof, a homolog thereof, a derivative thereof, or combinations thereof. Further embodiments pertain to methods of growing a modified plant or seed of the present disclosure in a field.

A fungal cell is capable of producing high levels of fatty acids and fatty acid-derived products. The fungal cell comprises at least one modification to the endogenous fatty acid metabolism.

The present disclosure provides recombinant microorganisms and methods for the anaerobic production of 2,4-furandicarboxylic acid from one or more carbon sources. The microorganisms and methods provide redox-balanced and ATP positive pathways for co-producing 2,4-furandicarboxylic acid with ethanol and for co-producing 2,4-furandicarboxylic acid with ethanol and 1-propanol. The method provides recombinant microorganisms that express endogenous and/or exogenous nucleic acid molecules encoding polypeptides that catalyze the conversion of a carbon source into 2,4-furandicarboxylic acid and that coupled the 2,4-furandicarboxylic acid pathway with an additional metabolic pathway.

The present invention relates, e.g., to in vitro method, using isolated protein reagents, for joining two double stranded (ds) DNA molecules of interest, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule share a region of sequence identity, comprising contacting the two DNA molecules in a reaction mixture with (a) a non-processive 5′ exonuclease; (b) a single stranded DNA binding protein (SSB) which accelerates nucleic acid annealing; (c) a non strand-displacing DNA polymerase; and (d) a ligase, under conditions effective to join the two DNA molecules to form an intact double stranded DNA molecule, in which a single copy of the region of sequence identity is retained. The method allows the joining of a number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes.

The present invention relates to enzyme inhibitors. More specifically, the present invention relates to ligand-directed covalent modification of proteins; method of designing same; pharmaceutical formulation of same; and method of use.