title: Generation of recombinant rabies viruses encoding NanoLuc luciferase for antiviral activity assays.
abstract: Rabies is an invariably fatal disease caused by Rabies virus (RABV), a member of the family Rhabdoviridae, genus Lyssavirus. Once central nervous infection occurs and symptoms develop, the case fatality rate approaches 100% despite availability of post-exposure prophylaxis. Therefore, new antiviral therapies for rabies are urgently required. Antivirals which can inhibit virus replication can be identified through screening of small compounds, however, as RABV infection does not generate easily discernible cytopathic effects in vitro, cell viability assays may not be feasible to observe antiviral activity of small compounds against RABV. In this study, recombinant RABVs (rRABVs) encoding NanoLuc luciferase (NanoLuc) were generated to facilitate the screening of small compound libraries. NanoLuc expression was confirmed in single-step growth cures of virus infection and showed that the rRABVs were capable of viral replication without decrease of luciferase activity through ten serial passages. Furthermore, the rRABVs were able to quantify the antiviral activity of the nucleoside analogue ribavirin against RABV in vitro. These findings confirm the potential of the rRABV encoding NanoLuc system to facilitate screening of small compounds to inhibit RABV infection.
authors: Division of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan.
authors: Paulina Duhita
authors: PD
authors: Anindita
authors: Division of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan.
authors: Michihito
authors: M
authors: Sasaki
authors: Division of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan; Discovery Research Laboratory for Core Therapeutic Areas, Shionogi & Co., Ltd., Osaka, Japan.
authors: Haruaki
authors: H
authors: Nobori
authors: Division of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan; Discovery Research Laboratory for Core Therapeutic Areas, Shionogi & Co., Ltd., Osaka, Japan.
authors: Akihiko
authors: A
authors: Sato
authors: Global Station for Zoonosis Control, Global Institution for Collaborative Research and Education (GI-CoRE), Hokkaido University, Sapporo, Japan; National Virus Reference Laboratory, University College Dublin, Belfield, Dublin 4, Ireland.
authors: Michael
authors: M
authors: Carr
authors: Laboratory of Zoonotic Diseases, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan.
authors: Naoto
authors: N
authors: Ito
authors: Laboratory of Zoonotic Diseases, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan.
authors: Makoto
authors: M
authors: Sugiyama
authors: Division of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan.
authors: Yasuko
authors: Y
authors: Orba
authors: Division of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan; Global Station for Zoonosis Control, Global Institution for Collaborative Research and Education (GI-CoRE), Hokkaido University, Sapporo, Japan; Global Virus Network, Baltimore, MD, United States. Electronic address: h-sawa@czc.hokudai.ac.jp.
authors: Hirofumi
authors: H
authors: Sawa
mesh_terms: D000998:Antiviral Agents / Q000302:isolation & purification*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D017930:Genes, Reporter; D008156:Luciferases / Q000032:analysis* / Q000235:genetics; D011820:Rabies virus / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000998:Antiviral Agents; D008156:Luciferases
keywords: Antiviral assay; NanoLuc luciferase; Recombinant rabies virus