Methods of improving homologous recombination

Details Methods of improving homologous recombination Methods of improving homologous recombination

C - Chemistry, Metallurgy

12

N

C12N 15/63 (2006.01) C07K 14/47 (2006.01) C12N 15/90 (2006.01)

Patent

CA 2403113

The invention features a method of promoting an alteration at a selected site in a target DNA, e.g., in the chromosomal DNA of a cell. The method includes providing, at the site: (a) a double stranded DNA sequence which includes a selected DNA sequence; (b) an agent which enhances homologous recombination, e.g., a Rad52 protein or a functional fragment thereof; and (c) an agent which inhibits non-homologous end joining, e.g., an agent which inactivates Ku such as an anti-Ku antibody or a Ku-binding oligomer or polymer, and allowing the alteration to occur. The agent which inhibits non-homologous end joining, e.g., a Ku inactivating agent such as an anti-Ku antibody, is preferably provided locally. Component (a), (b), and (c) can be introduced together, which is preferred, or separately.

L'invention concerne un procédé permettant de favoriser une modification sur un site sélectionné dans un ADN cible, par exemple, l'ADN chromosomique d'une cellule. Le procédé consiste à fournir à un site: (a) une séquence d'ADN à double hélice comprenant une séquence d'ADN sélectionnée; (b) un agent améliorant la recombinaison homologue, par exemple, une protéine Rad52 ou un fragment fonctionnel de celle-ci; et (c) un agent inhibant un joint d'extrémité non-homologue, par exemple, un agent qui inactive Ku, tel qu'un anticorps anti-Ku ou un polymère ou un oligomère se fixant à Ku; puis à permettant le déclenchement de la modification. De préférence, l'agent inhibant le joint d'extrémité non homologue, par exemple un agent inactivant Ku, tel qu'un anticorps anti-Ku, est fourni localement. Les composants (a), (b), et (c) peuvent être introduits ensemble, de préférence, ou séparément.

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