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 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,
wherein the polypeptide binds to DNA.
2. The isolated 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 protein comprises an atypical RVD as shown in Tables 27A or 27B.
4. The polypeptide of claim 1 , wherein the C-cap polypeptide is less than approximately 230 amino acids in length.
5. The polypeptide of claim 1 , wherein the C-cap comprises a TALE repeat domain.
6. A fusion protein comprising the polypeptide of claim 1 and at least one functional domain.
7. The fusion protein of claim 6 , wherein the functional domain is a transcriptional activator or a transcriptional repressor.
8. The fusion protein of claim 7 , wherein the functional domain comprises a nuclease.
9. The fusion protein of claim 8 , wherein the nuclease comprises at least one cleavage domain or cleavage half-domain from a TypeIIS endonuclease.
10. A host cell comprising a polypeptide according to claim 1 .
11. A pharmaceutical composition comprising a polypeptide according to claim 1 .