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US 8,912,138 B2Granted
DNA-binding proteins and uses thereof
AssigneeSangamo BioSciences, Inc.
InventorsPhilip D. Gregory, Jeffrey C. Miller, David Paschon, Edward J. Rebar, Siyuan Tan, Fyodor Urnov, Lei Zhang
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Abstract
Disclosed herein are polypeptides, polynucleotides encoding, cells and organisms comprising novel DNA-binding domains, including TALE DNA-binding domains. Also disclosed are methods of using these novel DNA-binding domains for modulation of gene expression and/or genomic editing of endogenous cellular sequences.
Claims13
A method of modifying an endogenous gene in a region of interest in a cell, the method comprising:
- introducing into the cell a polynucleotide encoding a fusion protein, the fusion protein comprising
- (a) an isolated, non-naturally occurring DNA-binding polypeptide comprising:
- two or more TALE-repeat units, the TALE repeat units comprising a repeat variable di-residue (RVD);
- an N-cap polypeptide, wherein the N-cap comprises residues N+1 to N+121 of a TALE protein;
- and
- a C-cap polypeptide, wherein the C-cap polypeptide comprises a fragment of a full length C-terminus region of a TALE protein, the C-cap comprising residues C-1 to C-20 of the TALE protein, and
- (b) a functional domain,
- wherein the fusion protein binds to a target site in the endogenous gene and further wherein the endogenous gene is modified.
5 more elements
Dependencies are inferred from the claim text.