DNA-binding proteins and uses thereof
First Claim
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1. 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.
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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.
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13 Claims
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1. A method of modifying an endogenous gene in a region of interest in a cell, the method comprising:
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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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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Specification