Abstract: This invention pertains to mutant Cas9 nucleic acids and proteins for use in CRISPR/Cas endonuclease systems, and their methods of use. In particular, the invention pertains to an isolated mutant Cas9 protein, wherein the isolated mutant Cas9 protein is active in a CRISPR/Cas endonuclease system, wherein the CRISPR/Cas endonuclease system displays reduced off-target editing activity and maintained on-target editing activity relative to a wild-type CRISPR/Cas endonuclease system. The invention also includes isolated nucleic acids encoding mutant Cas9 proteins, ribonucleoprotein complexes and CRSPR/Cas endonuclease systems having mutant Cas9 proteins that display reduced off-target editing activity and maintained on-target editing activity relative to a wild-type CRISPR/Cas endonuclease system.

Abstract: This invention pertains to mutant Lachnospiraceae bacterium ND2006 (Lb) Cas12a nucleic acids and proteins for use in CRISPR/Cas12a endonuclease systems, and their methods of use. In particular, the invention pertains to an isolated mutant LbCas12a protein, wherein the isolated mutant LbCas12a protein is active in a CRISPR/Cas12a endonuclease system. The invention also includes isolated nucleic acids encoding mutant LbCas12a proteins, ribonucleoprotein complexes and CRISPR/Cas12a endonuclease systems having mutant LbCas12a proteins.

Abstract: Described herein is a system and process for identifying and characterizing double-stranded DNA break repair sites that are based on biological information and have improved accuracy. Also described is a sequence alignment process that uses biological data to inform the alignment matrix for position specific alignment scoring, resulting in the identification of noncanonical target sites.

Abstract: The present invention is directed to methods and compositions for adding tails of specific lengths to a substrate polynucleotide. The invention also contemplates methods and compositions for immobilization of tailed substrates to a solid support. The disclosure contemplates that the attenuator molecule is any biomolecule that associates with a tail sequence added to a substrate polynucleotide and controls the addition of a tail sequence to the 3? end of the substrate polynucleotide. The sequence that is added to the substrate polynucleotide is referred to herein as a tail sequence, or simply a tail, and the process of adding a nucleotide to a substrate polynucleotide is referred to herein as tailing.

Abstract: This invention pertains to mutant Cas9 nucleic acids and proteins for use in CRISPR/Cas endonuclease systems, and their methods of use. In particular, the invention pertains to an isolated mutant Cas9 protein, wherein the isolated mutant Cas9 protein is active in a CRISPR/Cas endonuclease system, wherein the CRISPR/Cas endonuclease system displays reduced off-target editing activity and maintained on-target editing activity relative to a wild-type CRISPR/Cas endonuclease system. The invention also includes isolated nucleic acids encoding mutant Cas9 proteins, ribonucleoprotein complexes and CRISPR/Cas endonuclease systems having mutant Cas9 proteins that display reduced off-target editing activity and maintained on-target editing activity relative to a wild-type CRISPR/Cas endonuclease system.

Abstract: This invention pertains to optimized protein fusion linkers for creating multi-functional chimeric proteins and methods of using the same. Additionally, the invention pertains to chimeric proteins for use in guided endonuclease systems.

Abstract: Described herein are methods and compositions for improved hybridization capture for enriching for target nucleic acid sequences from a population of nucleic acids. In one aspect, the methods and compositions are useful for Next Generation Sequencing (NGS) applications. The methods and compositions include combining in solution capture probes and target nucleic acid and permitting hybridization of the target nucleic acid to the capture probes under conditions to promote efficient hybridization. Following hybridization, the probe/target complex is immobilized to a capture material while simultaneously incubating at an optimum hybridization temperature. The incubation denatures unwanted nucleic acids from the capture probes further enriching for target nucleic acid.

Abstract: Methods for performing multiplex PCR-based enrichment of a target substrate are provided. Systems and methods for generating a sequencing library are also provided.

Abstract: Described herein are compositions and methods for improving homology directed repair (HDR) efficiency and reducing homology-independent integration following introduction of double strand breaks with engineered nucleases. Additionally, modifications to double stranded DNA donors to improve the donor potency and efficiency of homology directed repair following introduction of double stranded breaks with programmable nucleases.

Abstract: The present invention is directed to methods and compositions for adding tails of specific lengths to a substrate polynucleotide. The invention also contemplates methods and compositions for immobilization of tailed substrates to a solid support. The disclosure contemplates that the attenuator molecule is any biomolecule that associates with a tail sequence added to a substrate polynucleotide and controls the addition of a tail sequence to the 3? end of the substrate polynucleotide. The sequence that is added to the substrate polynucleotide is referred to herein as a tail sequence, or simply a tail, and the process of adding a nucleotide to a substrate polynucleotide is referred to herein as tailing.

Abstract: The present invention pertains to a kit for producing an extended primer, comprising at least one container providing a hybrid RNase H2 protein, including a hybrid RNase H2 protein comprising fragments of amino acid sequences from Pyrococcus abyssi (P. a.), Thermococcus kodakarensis (T. kod), and Pyrococcus furiosus organisms; a hybrid RNase H2 protein comprises amino acid residues 26-40 and residues 100-120 of T. kod RNase H2; a hybrid RNase H2 protein is selected from SEQ ID NO: 2 and 3; and a hybrid RNase H2 protein is selected from SEQ ID NO: 14-20.

Abstract: This invention pertains to modified compositions for use in CRISPR systems, and their methods of use. In particular, length-modified and chemically-modified forms of crRNA are described for use as a reconstituted guide RNA for interaction with Cas9 of CRISPR systems. The resultant length-modified and chemically-modified forms of crRNA are economical to produce and can be tailored to have unique properties relevant to their biochemical and biological activity in the context of the CRIPSR Cas9 endonuclease system.

Abstract: This invention pertains to isolated mutant Cas9 nucleic acids and proteins for use in CRISPR/Cas endonuclease systems and their methods of use. In particular, the invention pertains to an isolated mutant Cas9 protein, wherein the isolated mutant Cas9 protein is active in a CRISPR/Cas endonuclease system, wherein the CRISPR/Cas endonuclease system displays a lower ratio of reduced off-target editing activity to on-target editing activity for at least one target site relative to the corresponding ratio of reduced off-site editing activity to on-target editing activity of a wild-type CRISPR/Cas endonuclease system having a WT-Cas9 protein of SEQ ID NO:5.

Abstract: Chemical synthesizer systems and methods for operating the same. One method includes receiving a first queue of instructions including a plurality of delivery instructions for operating a delivery assembly with respect to a plurality of synthesis plates and a plurality of vacuum instructions, grouped in a plurality of vacuum sections, for operating a vacuum assembly with respect to the plurality of synthesis plates. The method also includes sequentially processing each instruction included in the first queue of instructions by (i) executing the instruction when the instruction is one of the plurality of delivery instructions and (ii) moving, when the instruction is one of the plurality of vacuum instructions, one of the plurality of vacuum sections including the instruction to a second queue of instructions and executing instructions included in the second queue of instruction in parallel with instructions included in the first queue of instructions.

Abstract: Described herein are compositions and methods for improving homology directed repair (HDR) efficiency and reducing homology-independent integration following introduction of double strand breaks with engineered nucleases. Additionally, modifications to double stranded DNA donors to improve the donor potency and efficiency of homology directed repair following introduction of double stranded breaks with programmable nucleases.

Abstract: The present invention pertains to hybrid RNase H2 proteins that include fragments of amino acid sequences from Pyrococcus abyssi (P.a.), Thermococcus kodakarensis (T.kod), and Pyrococcus furiosus organisms, as well as methods of using the same to improve mismatch discrimination and activity in a high-fidelity DNA polymerase buffer.

Abstract: Described herein are methods for the expression and purification of Cas13a and methods for detecting target RNA using Cas13a.

Abstract: Described herein are compositions and methods for the production and quantification of barcoded or unique molecular identifier (UMI)-labeled substrates. In one aspect, the substrate is a bead comprising a template oligonucleotide that is elongated by successive extension reactions to provide a bead with an oligonucleotide comprising a plurality of barcodes and conserved anchor regions. Methods are also described for quantifying the amount of template oligonucleotide loaded onto the substrate and the products of the extension reaction after each round and after the final extension.

Abstract: Described herein are CAS12A mutants from Lachnospiraceae bacterium and methods for use thereof. These mutants have enhanced DNA cleavage activities at non-canonical TTTT protospacer adjacent motifs (PAM) compared to the wild-type enzyme.

Abstract: Described herein are CAS12A mutants from Lachnospiraceae bacterium and methods for use thereof. These mutants have enhanced DNA cleavage activities at non-canonical TTTT protospacer adjacent motifs (PAM) compared to the wild-type enzyme.