Methods and kits for mass production of dsrna

Details Methods and kits for mass production of dsrna Methods and kits for mass production of dsrna

C - Chemistry, Metallurgy

12

P

C12P 19/34 (2006.01) C12N 7/00 (2006.01) C12N 15/11 (2006.01)

Patent

CA 2528252

The invention relates to methods and kits for propagating target nucleic acid in the form of double stranded RNA. This invention relates in particular to a method for mass production of dsRNA. The method comprises that a target nucleic acid is provided in a form replicable by an RNA-dependent RNA polymerase in a living cell, said replicable form of the target nucleic acid is contacted with said polymerase under conditions sufficient for template- directed RNA synthesis, wherein one of the reaction products is necessarily double-stranded (ds) RNA and said dsRNA products are recovered in a sufficiently pure form. The dsRNA products can be used in various applications, for example in gene silencing.

La présente invention concerne des procédés et des trousses permettant de propager un acide nucléique cible sous forme d'ARN double brin. L'invention concerne en particulier un procédé qui permet la production en série d'ARN double brin. Le procédé de l'invention consiste à obtenir un acide nucléique cible sous une forme réplicable par une ARN polymérase ARN dépendante dans une cellule vivante, à mettre ladite forme réplicable de l'acide nucléique cible en contact avec la polymérase dans des conditions suffisantes pour la synthèse d'ARN dirigée par matrice, l'un des produits de la réaction étant nécessairement un ARN double brin, et à récupérer lesdits produits ARN double brin sous une forme suffisamment pure. Les produits ARN double brin de l'invention peuvent être utilisés dans diverses applications, par exemple l'extinction génique.

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