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: B J Yoo, M J Selby, J Choe, B S Suh, S H Choi, J S Joh, G J Nuovo, H S Lee, M Houghton, J H Han
• Journal: Journal of virology (J Virol) Vol. 69 Issue 1 Pg. 32-8 (Jan 1995) ISSN: 0022-538X [Print] United States
• PMID: 7983724 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Culture Media, Serum-Free
• RNA, Viral

Topics

• Base Sequence
• Carcinoma, Hepatocellular
• Culture Media, Serum-Free
• Hepacivirus (genetics, pathogenicity)
• Humans
• Molecular Sequence Data
• RNA, Viral (genetics)
• Transcription, Genetic
• Transfection
• Tumor Cells, Cultured