Methods for large-scale production of recombinant aav vectors

C - Chemistry – Metallurgy – 12 – N

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C12N 15/86 (2006.01) C12N 5/10 (2006.01) C12N 7/01 (2006.01) C12N 15/864 (2006.01) C12N 15/869 (2006.01)

Patent

CA 2347129

Disclosed are HSV-1 amplicons that supply all necessary helper functions required for rAAV packaging and methods for their use. These HSV-1 amplicons have been shown to be capable of rescuing and replicating all forms of rAAV genomes including rAAV genomes introduced into cells by infection of rAAV virions, rAAV genomes transfected into cells on plasmids or proviral rAAV genomes integrated into cellular chromosomal DNA. Also provided are methods for preparing high-titer rAAV vector compositions suitable for gene therapy and the delivery of exogenous polynucleotides to selected host cells.

L'invention concerne des amplicons du virus de l'herpès HSV-1 qui remplissent toutes les fonctions d'assistance nécessaires à l'empaquetage du virus associé aux adénovirus recombinant (rAAV), et les méthodes d'utilisation correspondantes. Il a été démontré que ces amplicons HSV-1 peuvent sauvegarder et répliquer toutes formes de génomes rAAV, dont les génomes rAAV introduits dans les cellules par une infection par des virions rAAV, les génomes rAAV transfectés dans les cellules sur des plasmides, ou les génomes rAAV proviraux intégrés à l'ADN chromosomique cellulaire. L'invention concerne également des méthodes permettant de préparer des compositions de vecteurs rAAV à titre élevé, convenant pour la thérapie génique et l'apport de polynucléotides exogènes à des cellules hôtes sélectionnées.

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