title: Identification of host genes involved in hepatitis C virus replication by small interfering RNA technology.
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 (LTbeta), an adapter protein (TRAF2), 2 transcription factors (RelA and NFkappaB2), 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Global Pharmaceutical Research and Development, Antiviral Research, Abbott Laboratories, 200 Abbott Park Road, Abbott Park, IL 60064, USA. teresa.ng@abbott.com
authors.forename: Teresa I
authors.initials: TI
authors.lastname: Ng
authors.forename: Hongmei
authors.initials: H
authors.lastname: Mo
authors.forename: Tami
authors.initials: T
authors.lastname: Pilot-Matias
authors.forename: Yupeng
authors.initials: Y
authors.lastname: He
authors.forename: Gennadiy
authors.initials: G
authors.lastname: Koev
authors.forename: Preethi
authors.initials: P
authors.lastname: Krishnan
authors.forename: Rubina
authors.initials: R
authors.lastname: Mondal
authors.forename: Ron
authors.initials: R
authors.lastname: Pithawalla
authors.forename: Wenping
authors.initials: W
authors.lastname: He
authors.forename: Tanya
authors.initials: T
authors.lastname: Dekhtyar
authors.forename: Jeremy
authors.initials: J
authors.lastname: Packer
authors.forename: Mark
authors.initials: M
authors.lastname: Schurdak
authors.forename: Akhteruzzaman
authors.initials: A
authors.lastname: Molla
mesh_terms: D015195:Drug Design; D020869:Gene Expression Profiling / Q000379:methods*; D015967:Gene Expression Regulation, Viral / Q000235:genetics; D015723:Gene Library; D005820:Genetic Testing / Q000379:methods; D016174:Hepacivirus / Q000235:genetics* / Q000254:growth & development*; D019698:Hepatitis C, Chronic / Q000235:genetics / Q000821:virology*; D006801:Humans; D007372:Interferons / Q000276:immunology; D034741:RNA, Small Interfering; D012367:RNA, Viral / Q000235:genetics; D012093:Replicon / Q000235:genetics; D014779:Virus Replication / Q000235:genetics
chemical_list: D034741:RNA, Small Interfering; D012367:RNA, Viral; D007372:Interferons