Patent ID: 8748146

Claim:
An engineered chimeric endonuclease comprising: the nucleic acid binding domain of a nuclease deficient I-SceI linked to the nucleic acid cleavage domain of FokI, wherein the nucleic acid binding domain of I-SceI endonuclease comprises an amino acid substitution at position 44 and 145, wherein the wild type residue aspartic acid (D) at position 44 is replaced with asparagine (N) or alanine (A) and wherein the wild type residue aspartic acid (D) at position 145 is replaced with alanine (A), wherein the engineered chimeric endonuclease binds a recognition sequence motif recognized by I-SceI, and wherein the engineered chimeric endonuclease cleaves at a unique cleavage position outside of the I-SceI recognition sequence motif.