A Convenient and Biosafe Replicon with Accessory Genes of SARS-CoV-2 and Its Potential Application in Antiviral Drug Discovery
| العنوان: | A Convenient and Biosafe Replicon with Accessory Genes of SARS-CoV-2 and Its Potential Application in Antiviral Drug Discovery |
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| المؤلفون: | Liu Cao, Deyin Guo, Yanxi Ji, Wei-Chen Wu, Yun-Yun Jin, Liubing Du, Yanchun Xie, Jiang Yiling, Ji-An Pan, Fan Xing, Xiaoxue Peng, Fuxiang Zheng, Hanwen Lin, Mang Shi, Kuijie Tong |
| المصدر: | Virologica Sinica |
| بيانات النشر: | Springer Singapore, 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | 0301 basic medicine, medicine.drug_class, viruses, 030106 microbiology, Immunology, Biology, Virus Replication, Antiviral Agents, Virus, 03 medical and health sciences, Virology, medicine, Humans, Replicon, Gene, Subgenomic mRNA, SARS-CoV-2, RNA, COVID-19, Containment of Biohazards, Reverse genetics, Antiviral drug screening, 030104 developmental biology, Virion assembly, Molecular Medicine, Antiviral drug, Research Article |
| الوصف: | SARS-CoV-2 causes the pandemic of COVID-19 and no effective drugs for this disease are available thus far. Due to the high infectivity and pathogenicity of this virus, all studies on the live virus are strictly confined in the biosafety level 3 (BSL3) laboratory but this would hinder the basic research and antiviral drug development of SARS-CoV-2 because the BSL3 facility is not commonly available and the work in the containment is costly and laborious. In this study, we constructed a reverse genetics system of SARS-CoV-2 by assembling the viral cDNA in a bacterial artificial chromosome (BAC) vector with deletion of the spike (S) gene. Transfection of the cDNA into cells results in the production of an RNA replicon that keeps the capability of genome or subgenome replication but is deficient in virion assembly and infection due to the absence of S protein. Therefore, such a replicon system is not infectious and can be used in ordinary biological laboratories. We confirmed the efficient replication of the replicon by demonstrating the expression of the subgenomic RNAs which have similar profiles to the wild-type virus. By mutational analysis of nsp12 and nsp14, we showed that the RNA polymerase, exonuclease, and cap N7 methyltransferase play essential roles in genome replication and sgRNA production. We also created a SARS-CoV-2 replicon carrying a luciferase reporter gene and this system was validated by the inhibition assays with known anti-SARS-CoV-2 inhibitors. Thus, such a one-plasmid system is biosafe and convenient to use, which will benefit both fundamental research and development of antiviral drugs. Supplementary Information The online version contains supplementary material available at 10.1007/s12250-021-00385-9. |
| اللغة: | English |
| تدمد: | 1995-820X 1674-0769 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3dbe6a6822706b054a0c14370729061a http://europepmc.org/articles/PMC8127439 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....3dbe6a6822706b054a0c14370729061a |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 1995820X 16740769 |
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