Subgenomic replicon derived from a cell line infected with the hepatitis C virus

Recently, cell culture systems have been established, where a hepatitis C virus (HCV) subgenomic replicon was efficiently replicated and maintained for a long period. To see whether a HCV sequence derived from HCV-infected cultured cell sequence can be used for the construction of a functional repli...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2002-05, Vol.293 (3), p.993-999
Main Authors: Kishine, Hiroe, Sugiyama, Kazuo, Hijikata, Makoto, Kato, Nobuyuki, Takahashi, Hitoshi, Noshi, Takeshi, Nio, Yasunori, Hosaka, Masahiro, Miyanari, Yusuke, Shimotohno, Kunitada
Format: Article
Language:English
Subjects:
Cell Line
Clone Cells
Genome, Viral
Gentamicins - pharmacology
Hepacivirus - genetics
Hepacivirus - growth & development
Hepacivirus - metabolism
Hepatitis C virus
Huh-7
Humans
Mutation
Replication
Replicon
RNA, Viral - biosynthesis
RNA, Viral - genetics
Transfection
Viral Proteins - biosynthesis
Viral Proteins - genetics
Virus Replication
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Summary:Recently, cell culture systems have been established, where a hepatitis C virus (HCV) subgenomic replicon was efficiently replicated and maintained for a long period. To see whether a HCV sequence derived from HCV-infected cultured cell sequence can be used for the construction of a functional replicon, a HCV subgenomic RNA carrying a neomycin-resistant gene was constructed using the HCV genome RNA obtained from cultured cells infected with HCV. After transfection, G418-resistant Huh-7 cells were selected and subcloned. Finally, the production of HCV proteins and de novo synthesis of subgenomic RNA were confirmed in the selected cell clone, indicating that this subgenomic RNA replicated in cultured cells and functioned as a replicon. These results suggest that the HCV genome obtained from an in vitro HCV infection system with cultured cells can be used to develop a subgenomic replicon system with diverse HCV sequences.
ISSN:0006-291X
1090-2104
DOI:10.1016/S0006-291X(02)00342-X