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US 8,771,986 B2Granted
Engineered cleavage half-domains
AssigneeSangamo BioSciences, Inc.
InventorJeffrey C. Miller
Life of this patent
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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.
Claims18
A polynucleotide encoding a polypeptide comprising an engineered FokI cleavage half-domain, wherein
- (i) the engineered cleavage half-domain comprises a substitution 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, wherein residues 384 to 579 of wild-type Fold is shown in SEQ ID NO:1;
- (ii) the engineered cleavage half-domain fatins 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.
1 more element
Dependencies are inferred from the claim text.