Preparation of recombinant adenovirus carrying a rep gene of...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/86 (2006.01) C12N 5/10 (2006.01) C12N 15/861 (2006.01) C12N 15/864 (2006.01)

Patent

CA 2326847

Adenoviral particles are produced by incubating cells containing a helper adenovirus vector and a helper-dependent adenoviral vector including an Adeno- Associated Virus (AAV) rep gene, such as rep78. Cells are provided containing a helper adenovirus vector. A helper-dependent adenoviral vector including an AAV rep gene is introduced into the cells, for instance by infection with infective viral particles. The cells are incubated to produce adenoviral particles containing nucleic acid including the AAV rep gene. Advantageously, cells containing helper adenovirus vector are pre-incubated for 0.5-12 hours, preferably about 4 hours, to allow expression of viral proteins required for adenoviral genome replication before introducing the helper-dependent adenovirus vector including an AAV rep gene.

L'invention concerne la production de particules adénovirales par des cellules d'incubation contenant un vecteur d'adénovirus auxiliaire et un vecteur adénoviral dépendant du virus auxiliaire incluant un gène rep de virus adéno-associé (AAV), tel que rep78. L'invention concerne des cellules contenant un vecteur d'adénovirus auxiliaire. Un vecteur adénoviral dépendant du virus auxiliaire qui inclut un gène rep d'AAV est introduit dans les cellules, par exemple par infection à l'aide de particules virales infectieuses. Les cellules sont incubées de manière à produire des particules adénovirales contenant un acide nucléique qui inclut le gène rep d'AAV. Il est avantageux de préincuber les cellules contenant le vecteur d'adénovirus auxiliaire pendant 0,5-12 heures, de préférence environ 4 heures, pour permettre l'expression de protéines virales nécessaires à la réplication du génome adénoviral avant d'introduire le vecteur d'adénovirus dépendant du virus auxiliaire qui inclut un gène rep d'AAV.

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