title: Novel reporter system to monitor early stages of the hepatitis B virus life cycle.
abstract: A recombinant hepatitis B virus (HBV) expressing NanoLuc (NL) (HBV/NL) was produced by cotransfecting a plasmid containing a 1.2-fold HBV genome carrying the NL gene with a plasmid bearing a packaging-defective 1.2-fold HBV genome into a human hepatoma cell line, HepG2. We found that NL activity in HBV/NL-infected primary hepatocytes or sodium taurocholate cotransporting polypeptide-transduced human hepatocyte-derived cell lines increased linearly for several days after infection and was concordant with HBV RNA levels in the cells. Treatment of the virus-infected cells with HBV inhibitors reduced NL activity in a dose-dependent manner. Detection of HBV/NL infection, monitored by NL activity, was highly sensitive and less expensive than detection using the conventional method to evaluate HBV infection. In addition, because we also studied host factors, this system is applicable not only for studying the HBV life cycle, but also for exploring agent(s) that regulate HBV proliferation.
authors: Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Ichikawa, Japan.
authors: Hironori
authors: H
authors: Nishitsuji
authors: Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Ichikawa, Japan.
authors: Saneyuki
authors: S
authors: Ujino
authors: Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Ichikawa, Japan.
authors: Yuko
authors: Y
authors: Shimizu
authors: Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Ichikawa, Japan.|Central Pharmaceutical Research Institute, Japan Tobacco Inc., Osaka, Japan.
authors: Keisuke
authors: K
authors: Harada
authors: Research and Development Center, FUSO Pharmaceutical Industries, Ltd, Osaka, Japan.
authors: Jing
authors: J
authors: Zhang
authors: Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Ichikawa, Japan.
authors: Masaya
authors: M
authors: Sugiyama
authors: Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Ichikawa, Japan.
authors: Masashi
authors: M
authors: Mizokami
authors: Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Ichikawa, Japan.
authors: Kunitada
authors: K
authors: Shimotohno
mesh_terms: D055786:Gene Knockout Techniques; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000254:growth & development*; D006801:Humans; D008018:Life Cycle Stages; D008156:Luciferases / Q000235:genetics; D016133:Polymerase Chain Reaction; D034741:RNA, Small Interfering; D014162:Transfection; D014773:Virology / Q000379:methods*
chemical_list: D034741:RNA, Small Interfering; D008156:Luciferases
keywords: HBV; host factors; luciferase; reporter HBV; virus entry