Integrated Dna Technologies, Inc.

Christopher Anthony Vakulskas

Michael Allen Collingwood

Mark Aaron Behlke

Garrett Richard Rettig

Liyang Zhang

Nicole Mary Bode

Sarah Franz Beaudoin

Kristin Beltz

Rolf Turk

Nathaniel Hunter Roberts

Steve Ehren Glenn

The present disclosure concerns polynucleotides and amino acids of. Cas12a (Cpf1) and methods for their use for genome editing in eukaryotic cells.

Methods using novel CPF1 mutations that enhance the DNA cleavage activity ofsp. CPF1

The disclosure provides Moloney murine leukemia virus (MMLV) reverse transcriptase (RTase) mutants. The disclosure as provides suitable amino acid positions in MMLV RTase for mutagenesis and methods and kits for using MMLV RTase mutants to synthesize cDNA from RNA templates.

Reverse transcriptase mutants with increased activity and thermostability

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.

sp. Cas12a mutants with enhanced cleavage activity at non-canonical TTTT protospacer adjacent motifs

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.

S. pyogenes CAS9 mutant genes and polypeptides encoded by same

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 increased 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 increased on-target editing activity relative to a wild-type CRISPR/Cas endonuclease system.

Mutations inCas9 discovered by broad scanning mutagenesis demonstrate enhancement of DNA cleavage activity

This invention pertains to mutantND2006 (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.

ND2006 CAS12A mutant genes and polypeptides encoded by same

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.

CAS9 mutant genes and polypeptides encoded by same

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.

Optimized protein fusions and linkers

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

Expression and purification of Cas enzymes

Described herein are CAS12A mutants fromand 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.

sp. CAS12A mutants with enhanced cleavage activity at non-canonical TTTT protospacer adjacent motifs

Cas9 mutant genes and polypeptides encoded by same

The present disclosure concerns polynucleotides and amino acids ofsp. Cas12a (Cpf1) and methods for their use for genome editing in eukaryotic cells.

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This invention pertains to recombinant AsCpf1 and LbCpf1 nucleic acids and polypeptides for use in CRISPR/Cpf1 endonuclease systems and mammalian cell lines encoding recombinant AsCpf1 or LbCpf1 polypeptides. The invention includes recombinant ribonucleoprotein complexes and CRSPR/Cpf1 endonuclease systems having a suitable AsCpf1 crRNA is selected from a length-truncated AsCpf1 crRNA, a chemically-modified AsCpf1 crRNA, or an AsCpf1 crRNA comprising both length truncations and chemical modifications. Methods of performing gene editing using these systems and reagents are also provided.

Growing community of inventors

North Liberty, IA, United States of America

Christopher Anthony Vakulskas