Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line.
An estimated 170 million persons worldwide are infected with hepatitis C virus (HCV), a major cause of chronic liver disease. Despite increasing knowledge of genome structure and individual viral proteins, studies on virus replication and pathogenesis have been hampered by the lack of reliable and efficient cell culture systems. A full-length consensus genome was cloned from viral RNA isolated from an infected human liver and used to construct subgenomic selectable replicons. Upon transfection into a human hepatoma cell line, these RNAs were found to replicate to high levels, permitting metabolic radiolabeling of viral RNA and proteins. This work defines the structure of HCV replicons functional in cell culture and provides the basis for a long-sought cellular system that should allow detailed molecular studies of HCV and the development of antiviral drugs.
Lohmann
V
V
Institute for Virology, Johannes-Gutenberg University Mainz, Obere Zahlbacher Strasse 67, 55131 Mainz, Germany.
Körner
F
F
Koch
J
J
Herian
U
U
Theilmann
L
L
Bartenschlager
R
R
D006528:Carcinoma, Hepatocellular; D003001:Cloning, Molecular; D004351:Drug Resistance; D016679:Genome, Viral; D005839:Gentamicins / Q000494:pharmacology; D016174:Hepacivirus / Q000235:genetics / Q000502:physiology*; D006526:Hepatitis C / Q000821:virology; D006801:Humans; D008113:Liver Neoplasms; D012367:RNA, Viral / Q000096:biosynthesis* / Q000235:genetics; D012093:Replicon; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000821:virology*; D017361:Viral Nonstructural Proteins / Q000032:analysis / Q000235:genetics; D014776:Virus Cultivation; D014779:Virus Replication; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; D012324:RNA-Dependent RNA Polymerase
D005839:Gentamicins; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C010680:antibiotic G 418; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase