Patent ID: 7888121

Claim:
A method for cleaving genomic cellular chromatin in a region of interest of an isolated cell, the method comprising: (a) selecting first and second nucleotide sequences in the region of interest; (b) engineering first and second zinc finger binding domains to bind to the first and second nucleotide sequences; (c) constitutively expressing a first fusion protein in a cell, the first fusion protein comprising the first engineered zinc finger binding domain and a first FokI cleavage half-domain; (d) constitutively expressing a second fusion protein in the cell, the second fusion protein comprising: (i) the second zinc finger binding domain; and (ii) a second Fold cleavage half-domain; wherein one of the cleavage half-domains comprises an alteration in amino acid 490 of the wild-type dimerization interface of the cleavage half-domain, the first fusion protein binds to the first nucleotide sequence, the second fusion protein binds to the second nucleotide sequence located between 2 and 50 nucleotides from the first nucleotide sequence, and the cellular chromatin is cleaved in the region of interest.