Method for enhancing the cleavage activity of i-crei derived meganucleases
AssigneeCellectis
InventorsSylvestre Grizot, Agnes Gouble, Christophe Perez-Michaut
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Abstract
A method for enhancing the cleavage activity of an I-CreI derived meganuclease, comprising the site-specific mutation of at least one amino acid residue which is selected in the group consisting of: the glycine at position 19, the phenylalanine at position 54, the phenylalanine at position 87, the serine at position 79, the valine at position 105 and the isoleucine at position 132 of I-CreI, and its application for the manufacturing of meganuclease cleaving a DNA target of interest, for use in genome therapy (treatment of genetic diseases) and genome engineering (making of transgenic animals, transgenic plants and recombinant cell lines).
Claims86
A method for enhancing the cleavage activity of an I-CreI derived meganuclease, wherein the method comprises the site-specific mutation of at least one amino acid residue selected from the group consisting of:
- the glycine at position 19 (G19), the phenylalanine at position 54 (F54), the phenylalanine at position 87 (F87), the serine at position 79 (S79), the valine at position 105 (V105) and the isoleucine at position 132 (I132) of I-CreI.
31-37. (canceled)
A method for making an I-CreI derived heterodimeric meganuclease substantially free of at least one of the two homodimers resulting from the association of each monomer of said heterodimeric meganuclease, comprising the co-expression of the two monomers of an I-CreI derived heterodimeric meganuclease in a cell, wherein one of the two monomers comprises the G19S mutation.
Dependencies are inferred from the claim text.