Dna transfection system for the generation of infectious...

C - Chemistry – Metallurgy – 12 – N

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Details

C12N 15/85 (2006.01) A61K 39/145 (2006.01) C07K 14/11 (2006.01) C12N 7/00 (2006.01) C12N 15/44 (2006.01) C12N 15/86 (2006.01)

Patent

CA 2406100

The present invention is based on the development of a dual promoter system (preferably a RNA pol I-pol II system) for the efficient intracellular synthesis of viral RNA. The resultant minimal plasmid-based system may be used to synthesize any RNA virus, preferably viruses with a negative single stranded RNA genome. The viral product of the system is produced when the plasmids of the system are introduced into a suitable host cell. One application of the system is production of attenuated, reassortant influenza viruses for use as antigens in vaccines. The reassortant viruses generated by cotransfection of plasmids may comprise genes encoding the surface glycoproteins hemagglutinin and neuraminidase from an influenza virus currently infecting the population and the internal genes from an attenuated influenza virus. An advantageous property of the present invention is its versatility; the system may be quickly and easily adapted to synthesize an attenuated version of any RNA virus. Attenuated or inactivated RNA viruses produced by the present invention may be administered to a patient in need of vaccination by any of several routes including intranasally or intramuscularly.

L'invention est basée sur l'élaboration d'un système à double promoteur (de préférence un système polI-polII d'ARN) servant à effectuer la synthèse intracellulaire d'ARN viral. On peut utiliser ce système basé sur un niveau minimum de plasmides afin de synthétiser tout virus d'ARN, de préférence, des virus possédant un génome négatif d'ARN monocaténaire. On obtient le produit viral de ce système quand on introduit ses plasmides dans une cellule hôte appropriée. Ce système trouve une de ses mises en application dans la production de virus atténués de réassortiment de la grippe conçus pour être utilisés en tant qu'antigènes dans des vaccins. Les virus de réassortiment générés par cotransfection de plasmide peuvent comprendre des gènes codant les glycoprotéines de surface hémagglutinine et neuraminidase provenant d'un virus de la grippe infectant fréquemment la population, et les gènes internes provenant d'un virus de la grippe atténué. Une propriété avantageuse de cette invention consiste en sa souplesse; on peut sans difficultés et rapidement adapter ce système à la synthèse d'une version atténuée de tout virus d'ARN. On peut administrer les virus d'ARN atténués ou inactivés, produits par l'invention, à un patient nécessitant une vaccination par toute voie possible, y compris la voie intranasale ou intramusculaire.

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