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...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.展开更多
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).
文摘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.
