Patent ID: 7972854

Claim:
A method for cleaving cellular chromatin in a region of interest, the method comprising: (a) selecting the region of interest; (b) engineering a first zinc finger binding domain to bind to a first nucleotide sequence in the region of interest; (c) providing a second zinc finger binding domain which binds to a second nucleotide sequence in the region of interest, wherein the second sequence is located between 2 and 50nucleotides from the first sequence; (d) expressing a first fusion protein in the cell, the first fusion protein comprising the first zinc finger binding domain and a first cleavage half-domain; and (e) expressing a second fusion protein in the cell, the second fusion protein comprising the second zinc finger binding domain and a second cleavage half domain; wherein (i) the first fusion protein binds to the first nucleotide sequence, (ii) the second fusion protein binds to the second nucleotide sequence, (iii) said binding of the first and second fusion proteins positions the cleavage half-domains such that the cellular chromatin is cleaved in the region of interest, and (iv) in at least one of the first or second fusion proteins, the cleavage half-domain is closer to the N-terminus and the zinc finger binding domain is closer to the C-terminus.