Accessory functions for use in recombinant aav virion...

C - Chemistry – Metallurgy – 12 – N

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

Patent

CA 2236968

Accessory functions capable of supporting efficient recombinant AAV (rAAV) virion production in a suitable host cell are provided. The accessory functions are in the form of one or more vectors comprising a nucleotide sequence selected from the group consisting of (i) a sequence that provides adenovirus VA RNAs, (ii) an adenovirus E4 ORF6 coding region, (iii) an adenovirus E2a 72 kD coding region, and any combination of nucleotide sequences (i), (ii) and (iii), and that are capable of being transferred between cells. Methods of producing rAAV virions are also provided. The methods can be practised to produce commercially significant levels of rAAV particles without also generating significant levels of infectious helper virus or other contaminating by-products.

On décrit des fonctions accesssoires qui permettent une production rentable de virions de virus adéno-associés (VAA) recombinés chez une cellule hôte appropriée. Elles prennent la forme d'un ou plusieurs vecteurs comprenant une séquence nucléotidique choisie dans le groupe consistant en: 1) une séquence qui donne des ARN VA d'adénovirus; 2) une région de codage E4 ORF 6 d'adénovirus; 3) une région de codage E2a de 72 dK d'adénovirus, ou l'une quelconque des combinaisons de ces séquences 1), 2) et 3), et qui peuvent être transférées entre des cellules. On décrit aussi des procédés de production de ces virions de VAA recombinés, utilisables pour produire des niveaux commercialement significatifs de particules de VAA recombinés sans qu'ils s'accompagnent de niveaux significatifs de virus auxiliaires infectieux ou d'autres sous-produits contaminants.

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