Methods and vectors for site-specific recombination

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/86 (2006.01) A61K 31/70 (2006.01) A61K 48/00 (2006.01) C12N 9/24 (2006.01) C12N 15/90 (2006.01)

Patent

CA 2230384

The present invention provides methods for site-specific recombination in a cell, as well as vectors which can be employed in such methods. The methods and vectors of the present invention can be used to obtain persistent gene expression in a cell and to modulate gene expression. One preferred method according to the invention comprises contacting a cell with a vector comprising an origin of replication functional in mammalian cells located between first and second recombining sites located in parallel. Another preferred method comprises, in part, contacting a cell with a vector comprising first and second recombining sites in antiparallel orientations such that the vector is internalized by the cell. In both methods, the cell is further provided with a site-specific recombinase that effects recombination between the first and second recombining sites of the vector.

L'invention concerne des procédés permettant une recombinaison dirigée dans une cellule, ainsi que des vecteurs utilisables dans ces procédés. Les procédés et les vecteurs de l'invention permettent d'obtenir une expression persistante des gènes dans une cellule et de moduler cette expression. Un procédé préféré selon l'invention consiste à mettre en contact une cellule avec un vecteur comprenant une origine de réplication, fonctionnant dans une cellule de mammifère, se trouvant entre les premier et second sites de recombinaison ayant des orientations parallèles. Un autre procédé préféré consiste à mettre en contact une cellule avec un vecteur comprenant des premier et second sites de recombinaison avec des orientations antiparallèles, de sorte que le vecteur subit une internalisation dans la cellule. Dans les deux procédés, la cellule est en outre pourvue d'une recombinase à action dirigée permettant une recombinaison entre les premier et second sites de recombinaison du vecteur.

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