An optimized high-throughput SARS-CoV-2 dual reporter trans-complementation system for antiviral screening <i>in vitro</i> and <i>in vivo</i>

Yingjian Li; Xue Tan; Jikai Deng; Xuemei Liu; Qianyun Liu; Zhen Zhang; Xiaoya Huang; Chao Shen; Ke Xu; Li Zhou; Yu Chen

doi:10.1016/j.virs.2024.03.009

June 2024

Citation: Yingjian Li, Xue Tan, Jikai Deng, Xuemei Liu, Qianyun Liu, Zhen Zhang, Xiaoya Huang, Chao Shen, Ke Xu, Li Zhou, Yu Chen. An optimized high-throughput SARS-CoV-2 dual reporter trans-complementation system for antiviral screening in vitro and in vivo .VIROLOGICA SINICA, 2024, 39(3) : 447-458. http://dx.doi.org/10.1016/j.virs.2024.03.009

An optimized high-throughput SARS-CoV-2 dual reporter trans-complementation system for antiviral screening in vitro and in vivo

a. State Key Laboratory of Virology, RNA Institute, College of Life Sciences and Frontier Science Center for Immunology and Metabolism, Wuhan University, Wuhan, 430072, China;
b. Institute for Vaccine Research at Animal Bio-safety Level Ⅲ Laboratory, Wuhan University School of Medicine, Wuhan, 430071, China

Corresponding author: Yu Chen, chenyu@whu.edu.cn
Received Date: 04 December 2023
Accepted Date: 21 March 2024
Available online: 26 March 2024

Abstract

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is still epidemic around the world. The manipulation of SARS-CoV-2 is restricted to biosafety level 3 laboratories (BSL-3). In this study, we developed a SARS-CoV-2 ΔN-GFP-HiBiT replicon delivery particles (RDPs) encoding a dual reporter gene, GFP-HiBiT, capable of producing both GFP signal and luciferase activities. Through optimal selection of the reporter gene, GFP-HiBiT demonstrated superior stability and convenience for antiviral evaluation. Additionally, we established a RDP infection mouse model by delivering the N gene into K18-hACE2 KI mouse through lentivirus. This mouse model supports RDP replication and can be utilized for in vivo antiviral evaluations. In summary, the RDP system serves as a valuable tool for efficient antiviral screening and studying the gene function of SARS-CoV-2. Importantly, this system can be manipulated in BSL-2 laboratories, decreasing the threshold of experimental requirements.
- SARS-CoV-2
- , RDP
- , Antiviral evaluation
- , Mouse model
- , BSL-2

Electronic Supplementary Material

10.1016j.virs.2024.03.009-ESM.docx
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