Engineered cleavage half-domains
First Claim
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1. A fusion protein comprising a polypeptide comprising an engineered FokI cleavage half-domain, wherein the engineered cleavage half-domain comprises a mutation selected from the group consisting of:
- substitution mutations in amino acid residues 486, 499 and 496;
substitution mutations in amino acid residues 487 and 496;
substitution mutations in amino acid residues 487, 499, and 496;
substitution mutations in amino acid residues 483 and 537;
substitution mutations in amino acid residues 490 and 537;
substitution mutations in amino acid residues 490, 537 and 538;
substitution mutations in amino acid residues 487, 499 and 496; and
substitution mutations in amino acid residues 483, 538 and 537, wherein the amino acid residues are numbered relative to full length wild-type FokI as shown in SEQ ID NO;
57.
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Abstract
Disclosed herein are engineered cleavage half-domains; fusion polypeptides comprising these engineered cleavage half-domains; polynucleotides encoding the engineered cleavage half-domains and fusion proteins; and cells comprising said polynucleotides and/or fusion proteins. Also described are methods of using these polypeptides and polynucleotides, for example for targeted cleavage of a genomic sequence.
42 Citations
21 Claims
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1. A fusion protein comprising a polypeptide comprising an engineered FokI cleavage half-domain, wherein the engineered cleavage half-domain comprises a mutation selected from the group consisting of:
- substitution mutations in amino acid residues 486, 499 and 496;
substitution mutations in amino acid residues 487 and 496;
substitution mutations in amino acid residues 487, 499, and 496;
substitution mutations in amino acid residues 483 and 537;
substitution mutations in amino acid residues 490 and 537;
substitution mutations in amino acid residues 490, 537 and 538;
substitution mutations in amino acid residues 487, 499 and 496; and
substitution mutations in amino acid residues 483, 538 and 537, wherein the amino acid residues are numbered relative to full length wild-type FokI as shown in SEQ ID NO;
57. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21)
- substitution mutations in amino acid residues 486, 499 and 496;
Specification