Several variants of concern(VOCs)have emerged since the WIV04 strain of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)was first isolated in January 2020.Due to mutations in the spike(S)protein,these VOCs have evolved to enhance viral infectivity and immune evasion.However,whether mutations of the other viral proteins lead to altered viral propagation and drug resistance remains obscure.The replicon is a nonin-fectious viral surrogate capable of recapitulating certain steps of the viral life cycle.Although several SARS-CoV-2 replicons have been developed,none of them were derived from emerging VOCs and could only recapitulate viral genome replication and subgenomic RNA(sgRNA)transcription.In this study,SARS-CoV-2 replicons derived from the WIV04 strain and two VOCs(the Beta and Delta variants)were prepared by removing the S gene from their genomes,while other structural genes remained untouched.These replicons not only recapitulate viral genome replication and sgRNA transcription but also support the assembly and release of viral-like particles,as manifested by electron microscopic assays.Thus,the S-deletion replicon could recapitulate virtually all the post-entry steps of the viral life cycle and provides a versatile tool for measuring viral intracellular propagation and screening novel antiviral drugs,including inhibitors of virion assembly and release.Through the quantification of replicon RNA released into the supernatant,we demonstrate that viral intracellular propagation and drug response to remde-sivir have not yet substantially changed during the evolution of SARS-CoV-2 from the WIV04 strain to the Beta and Delta VOCs.

Lingqian Tian;Qiuhong Liu;Rongjuan Pei;Yingshan Chen;Chonghui Xu;Jielin Tang;Hao Sun;Kunpeng Liu;Qi Yang;Lei Yang;Leshan Li;Yongli Zhang;Yuan Zhou;Chao Shan;Xue Hu;Xinwen Chen;Yun Wang

State Key Laboratory of Virology,Wuhan Institute of Virology,Center for Biosafety Mega-Science,Chinese Academy of Sciences,Wuhan,430071,China;University of Chinese Academy of Sciences,Beijing 100049,China;Guangzhou Institutes of Biomedicine and Health,Chinese Academy of Sciences,Guangzhou,510530,China;Department of Gastroenterology,Guangzhou Women and Children's Medical Center,Guangzhou,510623,China;Innovation Center for Pathogen Research,Guangzhou Laboratory,Guangzhou,510320,China

中国病毒学

[1]Lingqian Tian;Qiuhong Liu;Rongjuan Pei;Yingshan Chen;Chonghui Xu;Jielin Tang;Hao Sun;Kunpeng Liu;Qi Yang;Lei Yang;Leshan Li;Yongli Zhang;Yuan Zhou;Chao Shan;Xue Hu;Xinwen Chen;Yun Wang-.Comparison of viral propagation and drug response among SARS-CoV-2 VOCs using replicons capable of recapitulating virion assembly and release)[J].中国病毒学,2022(05):695-703

replicons,WIV04,replicon,nonin,fectious,subgenomic,remde,sivir

Comparison,propagation,response,among,SARS,CoV,VOCs,using,capable,recapitulating,virion,assembly,Several,variants,concern,have,emerged,since,strain,severe,acute,respiratory,syndrome,coronavirus,was,first,isolated,January,Due,mutations,spike,these,evolved,enhance,infectivity,immune,evasion,However,whether,other,proteins,lead,altered,resistance,remains,obscure,surrogate,certain,steps,life,cycle,Although,several,been,developed,none,them,were,derived,from,emerging,could,only,recapitulate,replication,sgRNA,transcription,In,this,study,two,Beta,Delta,prepared,by,removing,their,genomes,while,structural,genes,remained,untouched,These,not,but,also,support,like,particles,manifested,electron,microscopic,assays,Thus,deletion,virtually,post,entry,provides,versatile,tool,measuring,intracellular,screening,novel,antiviral,drugs,including,inhibitors,Through,quantification,released,into,supernatant,demonstrate,that,yet,substantially,changed,during,evolution

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