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AssigneesHvidovre Hospital, Kobenhavns Universitet
InventorsJudith M. Gottwein, Troels Kasper Hoyer Scheel, Jens Bukh, Jannick Prento, Tanja Bertelsen Jensen, Jacob Bo Lademann, Yiping Li
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Hepatitis C reporter viruses containing Core through NS2 of prototype isolates of all major HCV genotypes and the remaining genes of isolate JFH1, by insertion of reporter genes in domain III of HCV NS5A were developed. A deletion upstream of the inserted reporter gene sequence conferred favorable growth kinetics in Huh7.5 cells to these viruses. These reporter viruses can be used for high throughput analysis of drug and vaccine candidates as well as patient samples. JFH1-based intergenotypic recombinants with genotype specific homotypic 5′UTR, or heterotypic 5′UTR (either of genotype 1a (strain H77) or of genotype 3a (strain S52)) were also developed. The present inventors additionally developed J6/JFH1 recombinants with the 5′UTR of genotypes 1-6. These recombinants with different 5′UTRs are a useful to study the function of the 5′UTR in a genotype specific manner.
A chimeric nucleic acid molecule comprising the structural genes Core, E1, and E2, the p7 gene, and the non-structural gene NS2 of any one of a genotype 1a, 1b, 2a, 2b, 3a, 4a, 5a, 6a, or 7a Hepatitis C virus (HCV) and the non-structural genes NS3, NS4A, NS4B, NS5A and NS5B from the HCV genotype 2a JFH1 strain, wherein a reporter gene is inserted in domain III of the non-structural gene NS5A, wherein said NS5A gene encodes a non-structural NS5A protein having a 40 amino acid deletion of amino acid positions 250 to 289 or having a 25 amino acid deletion of amino acid positions 266-290, and wherein said nucleic acid encodes an intragenotypic or intergenotypic recombinant HCV genome that is infective and genetically stable in cell cultures.
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