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

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

机构地区：[1]State Key Laboratory of Virology,RNA Institute,College of Life Sciences and Frontier Science Center for Immunology and Metabolism,Wuhan University,Wuhan,430072,China [2]Institute for Vaccine Research at Animal Bio-safety Level III Laboratory,Wuhan University School of Medicine,Wuhan,430071,China

出　　处：《Virologica Sinica》2024年第3期447-458,共12页中国病毒学（英文版）

基　　金：supported by grants from the National Key R&D Program of China(2021YFA1300801 and 2018YFA0900801);National Natural Science Foundation of China(82172243 and 82372223);Fundamental Research Funds for the Central Universities(2042021kf0220 and 2042022dx0003).

摘　　要：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-2ΔN-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.

关 键 词：SARS-CoV-2 RDP Antiviral evaluation Mouse model BSL-2 

分 类 号：R563.1[医药卫生—呼吸系统]