In vitro system for replication of rna-dependent rna...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/11 (2006.01) C07H 21/02 (2006.01) C07H 21/04 (2006.01) C12N 5/08 (2006.01) C12N 15/33 (2006.01) C12N 15/85 (2006.01) C12Q 1/02 (2006.01) C12Q 1/68 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2437072

An in vitro method to conduct genomic replication of the viral genomes of viruses that utilize RNA-dependent RNA polymerase for replication (RDRP viruses), such as HCV. The method employs a construct comprising the 3' and 5' untranslated regions (UTRs) of the viral genome which are operably linked on the 5' and 3' ends of a reporter sequence, in antisense orientation, such that when viral replication is occurring within the cell which produces RDRP, the reporter protein will be made. The method of the invention provides an efficient means for measuring genomic replication in RDRP viruses, and also for the rapid screening of compounds for their ability to inhibit genomic replication of RDRP viruses, including the Hepatitis C virus (HCV).

Procédé <i>in vitro</i> pour mettre en oeuvre une réplication génomique de génomes viraux de virus qui utilisent l'ARN polymérase ARN dépendante pour se répliquer (virus RDRP), p. ex. virus de l'hépatite C (VHC). Le procédé utilise un gène hybride comportant les régions 3' et 5' non traduites (UTR) du génome viral, qui sont liées de manière opérationnelle sur les extrémités 5' et 3' d'une séquence de rapporteur, dans une orientation antisens, de sorte que lorsque la réplication virale se produit dans la cellule produisant RDRP, la protéine de rapporteur sera produite. Ce procédé constitue un moyen efficace pour mesurer la réplication génomique dans des virus RDRP, et est aussi utile dans le criblage rapide de composés pour leur capacité à inhiber la réplication génomique de virus RDRP, y compris le VHC.

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