A high throughput antiviral screening platform for alphaviruses based on Semliki Forest virus expressing eGFP reporter gene

Yu-Jia Shi; Jia-Qi Li; Hong-Qing Zhang; Cheng-Lin Deng; Qin-Xuan Zhu; Bo Zhang; Xiao-Dan Li

doi:10.1016/j.virs.2023.06.007

August 2023

Citation: Yu-Jia Shi, Jia-Qi Li, Hong-Qing Zhang, Cheng-Lin Deng, Qin-Xuan Zhu, Bo Zhang, Xiao-Dan Li. A high throughput antiviral screening platform for alphaviruses based on Semliki Forest virus expressing eGFP reporter gene .VIROLOGICA SINICA, 2023, 38(4) : 585-594. http://dx.doi.org/10.1016/j.virs.2023.06.007

A high throughput antiviral screening platform for alphaviruses based on Semliki Forest virus expressing eGFP reporter gene

Yu-Jia Shi ^a ,
Jia-Qi Li ^b,c ,
Hong-Qing Zhang ^b,c ,
Cheng-Lin Deng ^b,c ,
Qin-Xuan Zhu ^a ,
Bo Zhang ^{b,c
,,} ,
Xiao-Dan Li ^{a
,,}

a. Hunan Normal University, School of Medicine, Changsha, 410081, China;
b. Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan, 430071, China;
c. University of Chinese Academy of Sciences, Beijing, 100049, China

Corresponding author: Bo Zhang, zhangbo@wh.iov.cn
Xiao-Dan Li, lxd@live.cn
Received Date: 03 February 2023
Accepted Date: 20 June 2023
Available online: 28 June 2023

Abstract

Alphaviruses, which contain a variety of mosquito-borne pathogens, are important pathogens of emerging/re-emerging infectious diseases and potential biological weapons. Currently, no specific antiviral drugs are available for the treatment of alphaviruses infection. For most highly pathogenic alphaviruses are classified as risk group-3 agents, the requirement of biosafety level 3 (BSL-3) facilities limits the live virus-based antiviral study. To facilitate the antiviral development of alphaviruses, we developed a high throughput screening (HTS) platform based on a recombinant Semliki Forest virus (SFV) which can be manipulated in BSL-2 laboratory. Using the reverse genetics approach, the recombinant SFV and SFV reporter virus expressing eGFP (SFV-eGFP) were successfully rescued. The SFV-eGFP reporter virus exhibited robust eGFP expression and remained relatively stable after four passages in BHK-21 cells. Using a broad-spectrum alphavirus inhibitor ribavirin, we demonstrated that the SFV-eGFP can be used as an effective tool for antiviral study. The SFV-eGFP reporter virus-based HTS assay in a 96-well format was then established and optimized with a robust Z′ score. A section of reference compounds that inhibit highly pathogenic alphaviruses were used to validate that the SFV-eGFP reporter virus-based HTS assay enables rapid screening of potent broad-spectrum inhibitors of alphaviruses. This assay provides a safe and convenient platform for antiviral study of alphaviruses.
- Alphavirus
- , Semliki Forest virus
- , Reporter virus
- , High throughput screening
- , Inhibitor

Electronic Supplementary Material

10.1016j.virs.2023.06.007-ESM.docx
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