DNA-BINDING PROTEINS AND USES THEREOF
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.
1 . An isolated polynucleotide encoding a non-naturally occurring TALE DNA-binding polypeptide comprising:
two or more TALE-repeat units, the TALE repeat units comprising a repeat variable di-residue (RVD); and wherein the N-terminal region lacks at least 152 amino acids of a full-length TALE protein and the C-terminal region comprises fragment of a full-length C-terminal TALE protein, and further wherein the polypeptide binds to DNA.
2 . The polynucleotide of claim 1 , wherein at least one TALE-repeat unit comprises an atypical repeat variable di-residue (RVD).
3 . The polynucleotide of claim 2 , wherein the polypeptide comprises an atypical RVD as shown in Tables 27A or 27B.
4 . The polynucleotide of claim 1 , wherein the N-terminal region of the polypeptide is truncated to residue N+134 to residue N+137.
5 . The polynucleotide of claim 1 , wherein the C-cap of the polypeptide comprises a TALE repeat domain.
6 . The polynucleotide of claim 1 , wherein the polypeptide further comprises a functional domain.
7 . The polynucleotide of claim 6 , wherein the functional domain is a transcriptional activator or a transcriptional repressor.
8 . The polynucleotide of claim 1 , wherein the functional domain comprises a nuclease domain.
9 . The polynucleotide of claim 8 , wherein the nuclease domain is an endonuclease domain.
10 . A cell comprising the polynucleotide of claim 1 .
11 . A pharmaceutical composition comprising the polynucleotide of claim 1 .
12 . A pharmaceutical composition comprising a cell according to claim 10 .
13 . An isolated polynucleotide encoding a non-naturally occurring TALE DNA-binding polypeptide comprising:
two or more TALE-repeat units, the TALE repeat units comprising a repeat variable di-residue (RVD); and wherein the N-terminal region comprises amino acids N+1 to N+121 of a full-length TALE protein and the C-terminal region comprises no more than C−20 to C+63 of a full length TALE protein, and further wherein the polypeptide binds to DNA.
14 . The polynucleotide of claim 12 , wherein at least one TALE-repeat unit comprises an atypical repeat variable di-residue (RVD).
15 . The polynucleotide of claim 14 , wherein the polypeptide comprises an atypical RVD as shown in Tables 27A or 27B.
16 . The polynucleotide of claim 13 , wherein the N-terminal region of the polypeptide is truncated to residue N+134 to residue N+137.
17 . The polynucleotide of claim 13 , wherein the C-cap of the polypeptide comprises a TALE repeat domain.
18 . The polynucleotide of claim 13 , wherein the polypeptide further comprises a functional domain.
19 . The polynucleotide of claim 18 , wherein the functional domain is a transcriptional activator or a transcriptional repressor.
20 . The polynucleotide of claim 13 , wherein the functional domain comprises a nuclease domain.
21 . The polynucleotide of claim 20 , wherein the nuclease domain is an endonuclease domain.
22 . A cell comprising the polynucleotide of claim 13 .
23 . A pharmaceutical composition comprising the polynucleotide of claim 13 .
24 . A pharmaceutical composition comprising a cell according to claim 22 .