DOI: 10.1099/vir.0.81334-0. Development and characterization of a transient-replication assay for the genotype 2a hepatitis C virus subgenomic replicon

Development and characterization of a transient-replication assay for the genotype 2a hepatitis C virus subgenomic replicon

10.1099/vir.0.81334-0

Crossref journal-article

Microbiology Society

Journal of General Virology (345)

Abstract

Dicistronic, subgenomic hepatitis C virus (HCV) replicons were constructed containing sequences from JFH1, a genotype 2a strain, that also incorporated the firefly luciferase gene under the control of the HCV internal ribosome entry site element. Luciferase activity in Huh-7 cell extracts containing in vitro-transcribed subgenomic JFH1 RNA was monitored over a 72 h period to examine early stages of HCV replication in the absence of any selective pressure. Enzyme activities produced by the replicon were almost 200-fold greater than those generated from corresponding genotype 1b replicons and correlated with an accumulation of NS5A protein and replicon RNA. Transient replication was sensitive to IFN treatment in a dose-dependent manner and, in addition to Huh-7 cells, the U2OS human osteosarcoma cell line supported efficient replication of the JFH1 replicon. Thus, this system based on JFH1 sequences offers improvements over prior genotype 1b replicons for quantitative measurement of viral RNA replication.

Bibliography

Targett-Adams, P., & McLauchlan, J. (2005). Development and characterization of a transient-replication assay for the genotype 2a hepatitis C virus subgenomic replicon. Journal of General Virology, 86(11), 3075â3080.

Authors 2

Paul Targett-Adams (first)
John McLauchlan (additional)

References 20 Referenced 74

10.1126/science.290.5498.1972 / Science / Efficient initiation of HCV RNA replication in cell culture by Blight (2000)
10.1073/pnas.212532699 / Proc Natl Acad Sci U S A / Mutations that permit efficient replication of hepatitis C virus RNA in Huh-7 cells prevent productive replication in chimpanzees by Bukh (2002)
10.1074/jbc.M311120200 / J Biol Chem / Genotype 2a hepatitis C virus subgenomic replicon can replicate in HepG2 and IMY-N9 cells by Date (2004)
10.1128/JVI.76.12.5974-5984.2002 / J Virol / Expression of hepatitis C virus proteins induces distinct membrane alterations including a candidate viral replication complex by Egger (2002)
10.1099/0022-1317-82-4-723 / J Gen Virol / Interferon- α inhibits hepatitis C virus subgenomic RNA replication by an MxA-independent pathway by Frese (2001)
10.1128/JVI.75.24.12047-12057.2001 / J Virol / Sequences in the 5′ nontranslated region of hepatitis C virus required for RNA replication by Friebe (2001)
10.1128/JVI.77.9.5487-5492.2003 / J Virol / Identification of the hepatitis C virus RNA replication complex in Huh-7 cells harboring subgenomic replicons by Gosert (2003)
10.1128/JVI.75.18.8516-8523.2001 / J Virol / Effect of alpha interferon on the hepatitis C virus replicon by Guo (2001)
10.1016/j.pep.2004.05.005 / Protein Expr Purif / Purification and characterization of hepatitis C virus non-structural protein 5A expressed in Escherichia coli by Huang (2004)
10.1002/jmv.1055 / J Med Virol / Sequence analysis of hepatitis C virus isolated from a fulminant hepatitis patient by Kato (2001)
10.1053/j.gastro.2003.09.023 / Gastroenterology / Efficient replication of the genotype 2a hepatitis C virus subgenomic replicon by Kato (2003)
10.1128/JVI.79.1.592-596.2005 / J Virol / Nonhepatic cell lines HeLa and 293 support efficient replication of the hepatitis C virus genotype 2a subgenomic replicon by Kato (2005)
10.1128/JVI.75.10.4614-4624.2001 / J Virol / Enhancement of hepatitis C virus RNA replication by cell culture-adaptive mutations by Krieger (2001)
10.1126/science.285.5424.110 / Science / Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line by Lohmann (1999)
10.1128/JVI.77.5.3007-3019.2003 / J Virol / Viral and cellular determinants of hepatitis C virus RNA replication in cell culture by Lohmann (2003)
10.1002/hep.510280125 / Hepatology / Continuous human cell lines inducibly expressing hepatitis C virus structural and nonstructural proteins by Moradpour (1998)
10.1038/nm1268 / Nat Med / Production of infectious hepatitis C virus in tissue culture from a cloned viral genome by Wakita (2005)
10.1128/JVI.77.7.3898-3912.2003 / J Virol / Alpha interferon induces distinct translational control programs to suppress hepatitis C virus RNA replication by Wang (2003)
10.1006/viro.2002.1652 / Virology / Subgenomic hepatitis C virus replicons inducing expression of a secreted enzymatic reporter protein by Yi (2002)
10.1073/pnas.0503596102 / Proc Natl Acad Sci U S A / Robust hepatitis C virus infection in vitro by Zhong (2005)

Dates

Type	When
Created	19 years, 10 months ago (Oct. 14, 2005, 12:12 a.m.)
Deposited	4 years, 10 months ago (Oct. 22, 2020, 6:21 a.m.)
Indexed	4 months, 1 week ago (April 23, 2025, 5:45 a.m.)
Issued	19 years, 10 months ago (Nov. 1, 2005)
Published	19 years, 10 months ago (Nov. 1, 2005)
Published Print	19 years, 10 months ago (Nov. 1, 2005)

Funders 0

None

BibTeX

@article{Targett_Adams_2005, title={Development and characterization of a transient-replication assay for the genotype 2a hepatitis C virus subgenomic replicon}, volume={86}, ISSN={1465-2099}, url={http://dx.doi.org/10.1099/vir.0.81334-0}, DOI={10.1099/vir.0.81334-0}, number={11}, journal={Journal of General Virology}, publisher={Microbiology Society}, author={Targett-Adams, Paul and McLauchlan, John}, year={2005}, month=nov, pages={3075–3080} }

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