Abstract
A robust and genetically stable cell culture system for Hepatitis C Virus (HCV) genotype 3a is provided. A genotype 3a/2a (S52/JFH1) recombinant containing the structural genes (Core, E1, E2), p7 and NS2 of strain S52 was constructed and characterized in Huh7.5 cells. S52/JFH1 and J6/JFH viruses passaged in cell culture had comparable growth kinetics and yielded similar peak HCV RNA titers and infectivity titers. Direct genome sequencing of cell culture derived S52/JFH1 viruses identified putative adaptive mutations in Core, E2, p7, NS3, and NS5A; clonal analysis revealed that all genomes analyzed exhibited different combinations of these mutations. Finally, viruses resulting from transfection with RNA transcripts of five S52/JFH1 recombinants containing these combinations of putative adaptive mutations performed as efficiently as J6/JFH viruses in Huh7.5 cells and were all genetically stable after viral passage.
Original language | English |
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IPC | A61K 39/29 (20060101); C12N 15/06 (20060101); C12Q 1/02 (20060101); C07H 21/00 (20060101); C12N 7/02 (20060101); A61K 39/12 (20060101) |
Patent number | WO2008/125117 |
Filing date | 23/10/2008 |
Country/Territory | Denmark |
Priority date | 11/04/2008 |
Priority number | PCT/DK2008/050083 |
Publication status | Published - 3 Feb 2015 |