Methods using cre-lox for production of recombinant...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/86 (2006.01) C07K 14/015 (2006.01) C12N 7/01 (2006.01) C12N 9/00 (2006.01) C12N 15/861 (2006.01) C12N 15/864 (2006.01) A61K 48/00 (2006.01)

Patent

CA 2264499

Methods for efficient production of recombinant AAV are described. In one aspect, three vectors are introduced into a host cell. A first vector directs expression of cre recombinase, a second vector contains a promoter, a spacer sequence flanked by loxP sites and rep/cap, and a third vector contains a minigene containing a transgene and regulatory sequences flanked by AAV ITRs. In another aspect, the host cell stably or inducibly expresses cre recombinase and two vectors carrying the other elements of the system are introduced into the host cell.

Procédés de production efficace de virus adéno-associés (AAV) de recombinaison. Selon un aspect, trois vecteurs sont introduits dans une cellule hôte. Un premier vecteur dirige l'expression de la recombinase cre, un deuxième vecteur contient un promoteur, une séquence d'espacement flanquée de sites loxP et des gènes rep/cap et un troisième vecteur contient un minigène comportant un transgène et des séquences de régulation flanquées de séquences répétitives de terminaison inversées de AAV. Selon un autre aspect, la cellule hôte exprime de manière stable ou induite la recombinase cre et deux vecteurs portant les autres éléments du système sont introduits dans la cellule hôte.

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