I-dmoi derivatives with enhanced activity at 37·c and use...

Details I-dmoi derivatives with enhanced activity at 37·c and use... I-dmoi derivatives with enhanced activity at 37·c and use...

C - Chemistry, Metallurgy

12

N

C12N 9/22 (2006.01) A01H 5/00 (2006.01) A01K 67/027 (2006.01) C12N 15/55 (2006.01) C12N 15/62 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2564513

I-DmoI derivatives with enhanced cleavage activity at 37~C, said mutant comprising a sequence of a mutant of a I-DmoI endonuclease or a chimeric 5 derivative thereof including at least the first I-DmoIdomain, said sequence comprising the substitution of at least : (i) one of the residues in positions 4, 20, 49, 52, 92, 94 and/or 95 of said first I-DmoIdomain, and/or (ii) one of the residues in positions 101, 102, and/or 109 of the linker or the beginning of the second domain of I-DmoI, if present. 10 Polynucleotide encoding said derivatives, cell, animal or plant comprising said polynucleotide and use thereof for isolating meganucleases with new DNA target specificity.

La présente invention a trait à des dérivés de I-<I>Dmo</I>I à activité améliorée à 37 ·C, ledit mutant comportant une séquence d'un mutant I-<I>Dmo</I>I endonucléase ou un dérivé chimérique de celui-ci comprenant le premier domaine I-<I>Dmo</I>I, ladite séquence comportant la substitution d'au moins: (i) un des résidus en positions 4, 20, 49, 52, 92, 94 et/ou 95 dudit premier domaine I-<I>Dmo</I>I, et/ou (ii) un des résidus en positions 101, 102, et/ou 109 du lieur ou le début du deuxième domaine I-<I>Dmo</I>I, le cas échéant. L'invention a également trait à un polynucléotide codant pour lesdits dérivés, une cellule, un animal ou une plante comportant ledit polynucléotide et son utilisation pour l'isolement de méganucléases avec une nouvelle spécificité cible d'ADN.

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