Patent ID: 8206965

Claim:
An isolated hybrid meganuclease comprising two domains, wherein the N-terminal domain is derived from the first domain of the I-DmoI LAGLIDADG endonuclease and the C-terminal domain is derived from the I-Cre I LAGLIDADG endonuclease, wherein each of said N-terminal and C-terminal domain comprises a αββαββα fold, wherein the two ββ hairpins connected by an α helix (second helix) that are folded in a four-stranded antiparallel beta-sheet and the downstream α helix (third helix) form the DNA binding moiety of said domain, and wherein the first α helix comprises a dodecapeptide motif, wherein the αββαββα fold of the N-terminal domain corresponds to the amino acid sequence from positions 8 to 92 of the I-DmoI sequence SEQ ID NO: 18 and comprises the dodecapeptide motif SEQ ID NO: 20 (LLGLIIGDG), wherein the αββαββα fold of the C-terminal domain corresponds to the amino acid sequence from anyone of position 8 to 13 to position 94 of the I-CreI sequence SEQ ID NO: 19 and comprises the dodecapeptide motif SEQ ID NO: 21 (LAGFVDGDG), wherein said N-terminal and C-terminal domains are joined by a polypeptide linker, and wherein said hybrid meganuclease comprises an amino acid sequence which is at least 80% identical to SEQ ID NO: 2, and has endonuclease activity.