Adenovirus mutants with deleted protease gene, complementing...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/861 (2006.01) A61K 48/00 (2006.01) C12N 5/10 (2006.01) C12N 7/01 (2006.01)

Patent

CA 2322222

An adenovirus vector/packaging cell line system is disclosed, in which the vector replication is blocked by deletion of a single gene, which deletion does not interfere with any other viral functions. The deleted gene is the gene of the adenovirus protease. The protease is expressed in a complementing (packaging) cell line through a regulatable expression cassette which induces no toxic effects in the cells, thus making the generation and propagation of the vector easier and more efficient. As the deleted gene is highly specific of adenovirus, no complementation of the gene in transduced cells is expected, which increases the safety of the vectors for gene transfer purposes. Also disclosed is a system of generating recombinant adenovirus vectors by positive selection of recombinants deleted for the endogenous protease, gene which gene is cloned in another region of the adenoviral genome.

L'invention concerne un système de vecteur d'adénovirus/de lignée cellulaire d'encapsidation dans lequel la réplication du vecteur est bloquée par délétion d'un gène unique, laquelle délétion n'interfère avec aucune autre fonction virale. Le gène délété est celui de la protéase de l'adénovirus. La protéase est exprimée dans une lignée cellulaire de complémentation (encapsidation) par le biais d'une cassette d'expression régulable n'induisant pas d'effet toxique dans les cellules, rendant ainsi la reproduction et la propagation du vecteur plus facile et plus efficace. Etant donné que le gène délété est hautement spécifique de l'adénovirus, il n'y a aucune complémentation du gène dans les cellules transduites, ce qui augmente la sécurité des vecteurs destinés au transfert de gènes. L'invention concerne également un système de production de vecteurs d'adénovirus recombinés par sélection positive de recombinants délétés pour le gène de protéase endogène, lequel gène est cloné dans une autre région du génome adénoviral.

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