Generation of recombinant rabies viruses encoding NanoLuc luciferase for antiviral activity assays

doi:10.1016/j.virusres.2016.02.002

. 2016 Apr 2:215:121-8.

doi: 10.1016/j.virusres.2016.02.002. Epub 2016 Feb 8.

Generation of recombinant rabies viruses encoding NanoLuc luciferase for antiviral activity assays

Paulina Duhita Anindita¹, Michihito Sasaki¹, Haruaki Nobori², Akihiko Sato², Michael Carr³, Naoto Ito⁴, Makoto Sugiyama⁴, Yasuko Orba¹, Hirofumi Sawa⁵

Affiliations

¹ Division of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan.
² 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.
³ 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.
⁴ Laboratory of Zoonotic Diseases, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan.
⁵ 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.

PMID: 26869397
DOI: 10.1016/j.virusres.2016.02.002

Generation of recombinant rabies viruses encoding NanoLuc luciferase for antiviral activity assays

Paulina Duhita Anindita et al. Virus Res. 2016.

. 2016 Apr 2:215:121-8.

doi: 10.1016/j.virusres.2016.02.002. Epub 2016 Feb 8.

Authors

Paulina Duhita Anindita¹, Michihito Sasaki¹, Haruaki Nobori², Akihiko Sato², Michael Carr³, Naoto Ito⁴, Makoto Sugiyama⁴, Yasuko Orba¹, Hirofumi Sawa⁵

Affiliations

¹ Division of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan.
² 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.
³ 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.
⁴ Laboratory of Zoonotic Diseases, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan.
⁵ 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.

PMID: 26869397
DOI: 10.1016/j.virusres.2016.02.002

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.

Keywords: Antiviral assay; NanoLuc luciferase; Recombinant rabies virus.

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Generation of recombinant rabies viruses encoding NanoLuc luciferase for antiviral activity assays

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Generation of recombinant rabies viruses encoding NanoLuc luciferase for antiviral activity assays

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