CRISPR/CPF1 Systems and Methods
AssigneeIntegrated Dna Technologies, Inc.
InventorsMark Aaron BEHLKE, Michael Allen COLLINGWOOD, Rolf TURK, Christopher Anthony VAKULSKAS
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Abstract
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.
Claims23
1-24. (canceled)
An isolated nucleic acid, wherein the isolated nucleic acid encodes an Lb Cpf1 polypeptide codon optimized for expression in H. sapiens.
An isolated polypeptide comprising a wild-type Lp Cpf1 protein.
An isolated expression vector encoding SEQ ID NO:17 or SEQ ID NO:396.
A host cell comprising an isolated expression vector encoding SEQ ID NO:17 or SEQ ID NO:396, wherein the isolated expression vector encoding SEQ ID NO:17 or SEQ ID NO:
- 396 is operably linked to a suitable promoter to permit expression of a polypeptide comprising SEQ ID NO:14 or SEQ ID NO:24, respectively.
(canceled)
(canceled)
An isolated CRISPR/Cpf1 endonuclease system, comprising:
- an Lb Cpf1 polypeptide, and
- a suitable Cpf1 crRNA.
An isolated CRISPR/Cpf1 endonuclease system, comprising:
- a human cell line expressing a Lb Cpf1 polypeptide and a suitable Cpf1 crRNA.
A method of performing gene editing, comprising:
- contacting a candidate editing target site locus with an active CRISPR/Cpf1 endonuclease system having a wild-type Lb Cpf1 polypeptide and a suitable Cpf1 crRNA.
Dependencies are inferred from the claim text.