Productive Homologous and Non-homologous Recombination of Hepatitis C Virus in Cell Culture

• DOI: 10.1371/journal.ppat.1003228
• Publication Date: 2013-03-28T21:04:16Z
• Authors: Troels K. H. Scheel; Andrea Galli; Yi-Ping Li; Lotte S. Mikkelsen; Judith M. Gottwein; Jens Bukh
• Keywords: Viral evolution

Genetic recombination is an important mechanism for increasing diversity of RNA viruses, and constitutes a viral escape to host immune responses treatment with antiviral compounds. Although rare, epidemiologically hepatitis C virus (HCV) recombinants have been reported. In addition, regulatory cytopathogenicity the related pestiviruses. Here we describe HCV in cell culture leading production infectious virus. Initially, hepatoma cells were co-transfected replicating JFH1ΔE1E2 genome (genotype 2a) lacking functional envelope genes strain J6 (2a), which has but does not replicate culture. After initial decrease number positive cells, infection spread after 13-36 days. Sequencing recovered viruses revealed non-homologous sequence from 5' end NS2-NS3 region followed by JFH1 Core 3' end. These carried duplicated up 2400 nucleotides. replication was required recombination, as observed most experiments even when two incompetent genomes co-transfected. Reverse genetic studies verified viability representative recombinants. serial passage, subsequent events reducing or eliminating some all Furthermore, found that inter-genotypic could occur, at lower frequency than intra-genotypic recombination. Productive attenuated depended on expression proteins tolerated sequence. general, no strong site specificity observed. Non-homologous cases, while few homologous identified. A better understanding help identification natural thereby lead improved therapy. Our findings suggest mechanisms occurrence patients.