Compositions and methods to promote homologous recombination...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/55 (2006.01) A61K 31/70 (2006.01) A61K 48/00 (2006.01) C12N 9/00 (2006.01) C12N 15/79 (2006.01) C12N 15/90 (2006.01)

Patent

CA 2210706

The invention concerns genes encoding recombinases that can be used to promote homologous recombination in eukaryotic cells. The application teaches methods by which a recombinase of one species can be used to isolate a homologous recombinase of a different species and methods to identify the isolated homologs. Recombinases from Ustilago maydis and Saccharomyces cerevisiae are specifically included in the invention. The invention encompasses the method of producing an isolated recombinase protein in a prokaryotic cell and recovering the product in an active form. The invention also encompasses a genetically engineered gene which encodes a non-naturally occurring recombinase that causes a greater rate of recombination than does the naturally occurring recombinases. The invention further encompasses the use of recombinase proteins and of recombinase genes to promote homologous recombination, including recombination between a host cell genome and a chimeric oligonucleotide, i.e., an oligonucleotide having both RNA and DNA bases.

Le présent brevet porte sur des gènes codant pour des recombinases qui peuvent être utilisées pour faciliter la recombinaison homologue dans des cellules eucaryotes. On indique les méthodes par lesquelles une recombinase d'une espèce peut être utilisée pour isoler une recombinase homologue d'une autre espèce et les méthodes pour identifier les homologues isolés. Les recombinases d'Ustilago maydis et de Saccharomyces cerevisiae sont spécifiquement incluses dans le brevet. Il comprend la méthode d'obtention d'une recombinase isolée dans une cellule procaryote et la récupération du produit sous forme active. Il comprend aussi un gène obtenu par génie génétique qui code pour une recombinase de synthèse qui provoque un taux de recombinaison plus élevé que les recombinases naturelles. Le brevet divulgue également l'utilisation des recombinases et de gènes de la recombinase pour faciliter la recombinaison homologue, notamment la recombinaison entre le génome d'une cellule hôte et un oligonucléotide chimérique, c'est-à-dire un oligonucléotide possédant des bases de l'ARN et de l'ADN.

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