Modified zinc finger binding proteins
Disclosed herein are compositions and method comprising non-canonical (e.g., non-C2H2) zinc finger proteins.
1. An isolated polynucleotide encoding a non-naturally-occurring zinc-finger binding protein comprising a non-canonical zinc finger component, wherein:
said non-canonical zinc finger component comprises the sequence X 3 —Cys-X 2-4 —Cys-X 12 —His-X 1-7 —Z—X 4 (SEQ ID NO:121), where X is any amino acid and Z is C, where residues 6 to 12 of X 12 are engineered to bind to a target nucleic acid sequence, and wherein X 1-7 is selected from the group consisting of SETG (residues 2-5 of SEQ ID NO:12), LKSLTP (residues 2-7 of SEQ ID NO:13), SEN, VRI, KRS, KRL, LRW, KRT, ITT, IKG and IGG.
2. The isolated polynucleotide of claim 1 , wherein the target sequence is in an animal cell.
3. The isolated polynucleotide of claim 2 , wherein the target sequence is in a human cell.
4. The isolated polynucleotide of claim 1 , wherein the target sequence is a promoter sequence.
5. The isolated polynucleotide of claim 1 , wherein the zinc finger binding protein comprises three zinc finger components.
6. The isolated polynucleotide of claim 1 , wherein the target sequence comprises about 9 to about 14 contiguous base pairs.
7. The isolated polynucleotide of claim 5 , wherein the third zinc finger component comprises a non-canonical zinc finger component.
8. An expression vector comprising the polynucleotide of claim 1 .
9. An isolated host cell comprising the polynucleotide of claim 1 .
10. An isolated polynucleotide according to claim 1 further encoding a functional domain.
11. The polynucleotide of claim 10 , wherein the functional domain is a repressive domain.
12. The polynucleotide of claim 11 , wherein the repressive domain is selected from the group consisting of KRAB, MBD-2B, v-ErbA, MBD3, TR and members of the DNMT family.
13. The polynucleotide of claim 10 , wherein the functional domain is an activation domain.
14. The polynucleotide of claim 13 , wherein the activation domain is selected from the group consisting of maize C1, VP16, p65 subunit of NF-kappa B, and VP64.
15. The polynucleotide of claim 10 , wherein the functional domain is an endonuclease.
16. An expression vector comprising the polynucleotide of claim 10 .
17. An isolated host cell comprising the polynucleotide of claim 10 .
18. A composition comprising a polynucleotide according to claim 10 and a pharmaceutically acceptable excipient.
19. The isolated polynucleotide of claim 5 , wherein the first zinc finger component comprises a non-canonical zinc finger component.
20. The isolated polynucleotide of claim 1 , wherein the zinc finger binding protein comprises four zinc finger components.
21. The isolated polynucleotide of claim 1 wherein the target nucleotide sequence is in a plant cell.