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Identification of host genes involved in hepatitis C virus replication by small interfering RNA technology
Author(s) -
Ng Teresa I.,
Mo Hongmei,
PilotMatias Tami,
He Yupeng,
Koev Gennadiy,
Krishnan Preethi,
Mondal Rubina,
Pithawalla Ron,
He Wenping,
Dekhtyar Tanya,
Packer Jeremy,
Schurdak Mark,
Molla Akhteruzzaman
Publication year - 2007
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.21608
Subject(s) - ns5b , biology , small interfering rna , gene silencing , viral replication , rna interference , gene , hepatitis c virus , virology , subgenomic mrna , replicon , ns5a , gene knockdown , rna , virus , genetics , hepacivirus , plasmid
Abstract Hepatitis C virus (HCV) replication is highly dependent on host cell factors. Identification of these host factors not only facilitates understanding of the biology of HCV infection but also enables the discovery of novel targets for anti‐HCV therapy. To identify host genes important for HCV RNA replication, we screened a library of small interfering RNA (siRNA) that targets approximately 4,000 human genes in Huh7‐derived EN5‐3 cells harboring an HCV subgenomic replicon with the nonstructural region NS3‐NS5B from the 1b‐N strain. Nine cellular genes that potentially regulate HCV replication were identified in this screen. Silencing of these genes resulted in inhibition of HCV replication by more than 60% and exhibited minimal toxicity. Knockdown of host gene expression by these siRNAs was confirmed at the RNA level and, in some instances, at the protein level. The level of siRNA silencing of these host genes correlated well with inhibition of HCV. These genes included those that encoded a G‐protein coupled receptor ( TBXA2R ), a membrane protein ( LTβ ), an adapter protein ( TRAF2 ), 2 transcription factors ( RelA and NFκB2 ), 2 protein kinases ( MKK7 and SNARK ), and 2 closely related transporter proteins ( SLC12A4 and SLC12A5 ). Of interest, some of these genes are members of the tumor necrosis factor/lymphotoxin signaling pathway. Conclusion: Findings of this study may provide important information for understanding HCV replication. In addition, these cellular genes may constitute a novel set of targets for HCV antiviral therapy. (H EPATOLOGY 2007.)