title: Transfection of a differentiated human hepatoma cell line (Huh7) with in vitro-transcribed hepatitis C virus (HCV) RNA and establishment of a long-term culture persistently infected with HCV.
abstract: T7 RNA polymerase transcripts of a putative full-length cDNA clone of hepatitis C virus type 1 (HCV-1) were used to transfect a differentiated human hepatoma cell line, Huh7. The transfected genome replicated in cells, as evidenced by the appearance of progeny HCV RNA, detection of negative-strand viral RNA, and incorporation of [3H]uridine into the viral genome. Incubation of naive Huh7 cells with conditioned medium from transfected cells resulted in a new HCV infection, suggesting the production of biologically active virus in the inoculum. Maintenance of the transfected cells under serum-free culture conditions resulted in the selection of persistently infected cells which displayed a distinctive cellular morphology. This is the first demonstration that HCV RNA produced from cloned HCV cDNA is infectious and replication competent. This approach should provide a valuable system for studying HCV replication, persistence, and pathogenicity.
authors.affiliation: Chiron Corporation, Emeryville, California 94608.
authors.forename: B J
authors.initials: BJ
authors.lastname: Yoo
authors.forename: M J
authors.initials: MJ
authors.lastname: Selby
authors.forename: J
authors.initials: J
authors.lastname: Choe
authors.forename: B S
authors.initials: BS
authors.lastname: Suh
authors.forename: S H
authors.initials: SH
authors.lastname: Choi
authors.forename: J S
authors.initials: JS
authors.lastname: Joh
authors.forename: G J
authors.initials: GJ
authors.lastname: Nuovo
authors.forename: H S
authors.initials: HS
authors.lastname: Lee
authors.forename: M
authors.initials: M
authors.lastname: Houghton
authors.forename: J H
authors.initials: JH
authors.lastname: Han
mesh_terms: D001483:Base Sequence; D006528:Carcinoma, Hepatocellular; D016895:Culture Media, Serum-Free; D016174:Hepacivirus / Q000235:genetics* / Q000472:pathogenicity; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics*; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D016895:Culture Media, Serum-Free; D012367:RNA, Viral