Replication of Subgenomic Hepatitis C Virus RNAs in a Hepatoma Cell Line

TLDR

Hepatitis C virus infects ~170 million people worldwide and causes chronic liver disease, yet studies of its replication and pathogenesis have been limited by the lack of reliable cell culture systems. The study aims to define the structure of HCV replicons that function in cell culture, establishing a system for detailed molecular studies and antiviral drug development. The authors cloned a full-length consensus HCV genome from infected liver RNA and used it to construct subgenomic selectable replicons. The replicons replicated to high levels in a human hepatoma cell line, enabling metabolic radiolabeling of viral RNA and proteins and establishing a platform for detailed HCV studies and antiviral drug development.

Abstract

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.