Engineered cleavage half-domains
First Claim
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1. A polypeptide comprising an engineered FokI cleavage half-domain, wherein(i) the engineered cleavage half-domain comprises a mutation in one or more wild-type amino acid residue 483, 486, 487, 490, 499, 538 or combinations thereof, wherein the amino acid residues are numbered relative to full length wild-type FokI;
- (ii) the engineered cleavage half-domain forms an obligate heterodimer with a wild-type cleavage half-domain or a second engineered cleavage half-domain; and
(iii) if the engineered cleavage half-domain comprises a mutation at amino acid residue 490, it further comprises at least one additional amino acid mutation.
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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.
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Citations
15 Claims
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1. A polypeptide comprising an engineered FokI cleavage half-domain, wherein
(i) the engineered cleavage half-domain comprises a mutation in one or more wild-type amino acid residue 483, 486, 487, 490, 499, 538 or combinations thereof, wherein the amino acid residues are numbered relative to full length wild-type FokI; -
(ii) the engineered cleavage half-domain forms an obligate heterodimer with a wild-type cleavage half-domain or a second engineered cleavage half-domain; and (iii) if the engineered cleavage half-domain comprises a mutation at amino acid residue 490, it further comprises at least one additional amino acid mutation. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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Specification