Engineered zinc finger proteins for regulation of gene expression
Disclosed herein are methods and compositions for modulation of gene expression, with single-gene specificity, in a human-sized genome.
1. An isolated polynucleotide comprising a nucleic acid sequence encoding an engineered zinc finger protein, the engineered zinc finger protein comprising six zinc fingers, each zinc finger comprising a recognition region, wherein the amino acid sequence of the recognition regions, in amino- to carboxy-terminal order, are as follows: RSDHLSR (SEQ ID NO: 1); DNRDRTK (SEQ ID NO:2); DRKTLIE (SEQ ID NO:3); TSSGLSR (SEQ ID NO:4); RSDHLSE (SEQ ID NO:5); TSSDRTK (SEQ ID NO:6).
2. The polynucleotide of claim 1 , wherein the target site is in a chk2 gene.
3. The polynucleotide of claim 1 , further comprising a sequence encoding at least one functional domain.
4. The polynucleotide of claim 3 , wherein the functional domain comprises a transcriptional activation domain.
5. The polynucleotide of claim 3 , wherein the functional domain comprises a transcriptional repression domain.
6. The polynucleotide of claim 3 , wherein the functional domain comprises a nuclease domain.
7. An isolated polypeptide encoded by the polynucleotide of claim 1 .
8. A method of modulating expression of a chk2 gene in an isolated host cell; the method comprising administering a polynucleotide according to claim 3 to the host cell, wherein the polynucleotide expresses the zinc finger protein in the host cell and the zinc finger protein binds to the target site, thereby modulating the expression of the chk2 gene.
9. The method of claim 8 , wherein expression of the single chk2 gene is repressed.
10. The method of claim 8 , wherein expression of the single chk2 gene is activated.