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US 10,538,787 B2Granted
Compositions for linking DNA-binding domains and cleavage domains
AssigneeSangamo Therapeutics, Inc.
InventorsDavid Paschon, Lei Zhang
Life of this patent
Estimating the term…
Abstract
Disclosed herein are compositions for linking DNA binding domains and cleavage domains (or cleavage half-domains) to form non-naturally occurring nucleases. Also described are methods of making and using compositions comprising these linkers.
Claims16
A fusion protein comprising
- a DNA-binding domain having an N-terminus and a C-terminus, wherein the DNA-binding domain binds to a nucleotide target site;
- a truncated FokI cleavage domain in which the N-terminal residues QLVKS (residues 158 to 162 of SEQ ID NO:1) are deleted;
- and
- a linker between the C-terminus of the DNA-binding domain and the N-terminus of the cleavage domain, wherein the linker comprises a sequence selected from the group consisting of (N)GICPPPRPRTSPP (L8a, SEQ ID NO:2);
- (T)GTAPIEIPPEVYP (L8b, SEQ ID NO:3);
- (N)GSYAPMPPLALASP (L8c, SEQ ID NO:4);
- (P)GIYTAPTSRPTVPP (L8d, SEQ ID NO:5);
- (N)GSQTPKRFQPTHPSA (L8e, SEQ ID NO:6);
- TGLMPPSHPRQPIHINF (L8g, SEQ ID NO:234);
- TGTVHTSPICPQTYP (L8i, SEQ ID NO:161);
- TGSGTPTRPHPPLPP (L8j, SEQ ID NO:235);
- (H)LPKPANPFPLD (L7-1, SEQ ID NO:7);
- (H)RDGPRNLPPTSPP (L7-2, SEQ ID NO:8);
- (H)RLPDSPTALAPDTL (L7-6, SEQ ID NO:9);
- (D)PNSPISRARPLNPHP (L7-3, SEQ ID NO:10);
- (Y)GPRPTPRLRCPIDSLIFR (L7-5, SEQ ID NO:11);
- (H)CPASRPIHP (L6-2, SEQ ID NO:12);
- (G)LQSLIPQQLL (L6-6, SEQ ID NO:13);
- (G)LQPTVNHEYNN (L6-7, SEQ ID NO:14);
- and (P)ANIHSLSSPPPL (L6-1, SEQ ID NO:15) and further wherein the amino acid residue shown in round parentheses is optionally present.
17 more elements
Dependencies are inferred from the claim text.