Method of producing a recombinant adeno-associated virus,...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/864 (2006.01) A61K 48/00 (2006.01) C12N 5/10 (2006.01) C12N 15/19 (2006.01)

Patent

CA 2359748

The present invention relates to a method for producing a recombinant adeno-associated virus (rAAV), in which at different times a helper construct and also a vector construct are introduced into a suitable cell, and preferably the helper construct does not comprise, in particular with the exception of the AAV promoters, any nucleic acid sequences to which at least a Rep protein can essentially specifically bind and preferably the vector construct comprises ITR sequences in flop orientation. The recombinant adeno-associated viruses produced according to the method of the invention are suitable in particular for producing a tumor cell into which additionally nucleic acids coding for GM-CSF and B7.2 has been introduced, which in turn can be used in the form of a medicament for the treatment of cancers.

Cette invention concerne un procédé de fabrication d'un virus adéno-associé (AAV) recombiné. On introduit un assistant de recombinaison et un vecteur de recombinaison, de façon décalée dans le temps, à l'intérieur d'une cellule adaptée. Cet assistant ne contient de préférence, à l'exception, notamment, des promoteurs d'AAV, aucune séquence nucléotidique sur laquelle au moins une protéine Rep peut se lier de manière essentiellement spécifique. Le vecteur contient de préférence des séquences à répétitions terminales inversées orientées </= flop >/= . Les virus adéno-associés recombinés fabriqués au moyen du procédé selon l'invention conviennent particulièrement à la production d'une cellule tumorale dans laquelle on introduit des acides nucléiques supplémentaires codant pour le facteur stimulant les colonies de granulocytes macrophages ou "GM-CSF" ("Granulocyte/Macrophage Colony Stimulating Factor" ) et pour B7.2. Cette cellule peut à son tour être utilisée sous forme de médicament pour le traitement du cancer.

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