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Updated: Sep 26, 2026

A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
Identification of the hepatitis C virus RNA replication complex in Huh-7 cells harboring subgenomic replicons
Rainer Gosert1, Denise Egger, Volker Lohmann
1Department of Medicine II, University of Freiburg, Germany.
Abstract:
Formation of a membrane-associated replication complex, composed of viral proteins, replicating RNA, and altered cellular membranes, is a characteristic feature of plus-strand RNA viruses. Here, we demonstrate the presence of a specific membrane alteration, designated the membranous web, that contains hepatitis C virus (HCV) nonstructural proteins, as well as viral plus-strand RNA, in Huh-7 cells harboring autonomously replicating subgenomic HCV RNAs. Metabolic labeling with 5-bromouridine 5'-triphosphate in the presence of actinomycin D revealed that the membranous web is the site of viral RNA synthesis and therefore represents the replication complex of HCV.
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