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, 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 is truncated to residue C+63, and further wherein the polypeptide binds to DNA.
2. The polypeptide of claim 1 , wherein at least one TALE-repeat unit comprises an atypical repeat variable di-residue (RVD).
3. The polypeptide of claim 2 , wherein the polypeptide comprises an atypical RVD as shown in Tables 27A or 27B.
4. The polypeptide of claim 1 , wherein the N-terminal region is truncated to residue N+134 to residue N+137.
5. The polypeptide of claim 1 , wherein the C-cap comprises a TALE repeat domain.
6. The polypeptide of claim 1 , further comprising a functional domain.
7. The polypeptide of claim 6 , wherein the functional domain is a transcriptional activator or a transcriptional repressor.
8. The polypeptide of claim 1 , wherein the functional domain comprises a nuclease domain.
9. The polypeptide of claim 8 , wherein the nuclease domain is an endonuclease domain.
10. An isolated cell comprising the polypeptide of claim 1 .
11. A pharmaceutical composition comprising the cell according to claim 10 .
12. A pharmaceutical composition comprising the polypeptide according to claim 1 .