Drosophila recombination-associated protein and methods for use

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/52 (2006.01) A61K 48/00 (2006.01) C07K 16/40 (2006.01) C12N 9/00 (2006.01) C12N 15/10 (2006.01)

Patent

CA 2379802

The invention encompasses Drosophila Recombination Associated Protein (DRAP) isolated from D. melanogaster and a nucleic acid sequence encoding DRAP The Drosophila Recombination Associated Protein, its homologues from other organ- isms or active peptides derived therefrom, as well as DNA encoding such protein are useful for homology-dependent pairing of three DNA strands. The combination of strand-transfer and topoisomerase activities associated with DRAP permits directed pairing and cleavage at defined site(s) within DNA. This in turn makes possible the isolation and/or removal of a defined segment of DNA. DRAP is also useful in cloning, genomic cloning and gene mapping, in promoting gene disruptions or "knockout" mutations, in carrying out targeted mutagenesis of specific genes and in generating transgenic animals. The invention further encompasses a method for experimental and therapeutic application of DRAP driven knockouts or other modifications of genes responsible for genetic diseases as well as the use of DRAP driven genetic manipulation of genes in gene therapy.

L'invention concerne la protéine associée à la recombinaison de drosophila (DRAP) isolée de la <i>D. melanogaster</i> et une séquence d'acide nucléique codant pour la DRAP. Cette protéine, ses homologues provenant d'autres organismes ou de peptides actifs dérivés de ceux-ci, ainsi qu'un ADN codant pour une telle protéine sont utilisés pour appariement en fonction de l'homologie de trois brins d'ADN. La combinaison des activités brin-transfert et de topoisomérase associées à la DRAP permet un appariement et un clivage sur des sites définis contenus dans l'ADN. Ceci permet l'isolation et/ou le retrait d'un segment défini d'ADN. La DRAP est également utile pour le clonage, le clonage génomique et la cartographie génétique, pour favoriser la disruption génique ou les mutations de type inactivation, pour réaliser la mutagenèse ciblée de gènes spécifiques et pour créer des animaux transgéniques. L'invention comprend également un procédé permettant une application expérimentale et thérapeutique des inactivations à l'aide de la DRAP ou d'autres modifications de gènes responsables de maladies génétiques, ainsi que la manipulation génétique de gènes à l'aide de la DRAP en thérapie génique.

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