Patent ID: 8530214

Claim:
A polynucleotide encoding a first monomer of an I-CreI meganuclease variant said monomer comprising mutations in the amino acid sequence of SEQ ID NO: 34, wherein said mutations include: (i) at least one and up to eleven amino acid substitutions from residue S22 to residue Q44 said substitutions selected from the group consisting of substitutions at positions S22, I24, Q26, K28, N30, S32, Y33, Q38, S40, T42 and Q44, wherein the first monomer comprises amino acids from residue S22 to residue Q44 which correspond to SEQ ID NO: 34 wherein amino acids other than S22, I24, Q26, K28, N30, S32, Y33, Q38, S40, T42 and Q44 are not substituted; and (ii) at least one and up to six amino acid substitutions from residue Y66 to residue I77 said substitutions selected from the group consisting of substitutions at positions Y66, R68, R70, V73, D75 and I77, wherein the first monomer comprises amino acids from residue Y66 to residue I77 which correspond to SEQ ID NO: 34 wherein amino acids other than Y66, R68, R70, V73, D75 and I77 are not substituted; and wherein said monomer when in dimeric form binds and cleaves DNA.