{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T10:23:06Z","timestamp":1792146186671,"version":"4.4.0"},"update-to":[{"DOI":"10.1371\/journal.ppat.1009439","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2021,3,24]],"date-time":"2021-03-24T00:00:00Z","timestamp":1616544000000}}],"reference-count":45,"publisher":"Public Library of Science (PLoS)","issue":"3","license":[{"start":{"date-parts":[[2021,3,12]],"date-time":"2021-03-12T00:00:00Z","timestamp":1615507200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005089","name":"Beijing Municipal Natural Science Foundation","doi-asserted-by":"publisher","award":["M21001"],"award-info":[{"award-number":["M21001"]}],"id":[{"id":"10.13039\/501100005089","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Tsinghua University Initiative Scientific Research Program","award":["2019Z06QCX10"],"award-info":[{"award-number":["2019Z06QCX10"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32041005"],"award-info":[{"award-number":["32041005"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Key Research and Development Program of China","award":["2020YFA0707701"],"award-info":[{"award-number":["2020YFA0707701"]}]},{"name":"Tsinghua-Peking University Center of Life Sciences","award":["045-61020100120"],"award-info":[{"award-number":["045-61020100120"]}]},{"name":"Beijing Advanced Innovation Center for Structure Biology","award":["100300001"],"award-info":[{"award-number":["100300001"]}]},{"name":"Start-up Foundation of Tsinghua University","award":["53332101319"],"award-info":[{"award-number":["53332101319"]}]}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p>\n                    Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes the global pandemic of COVID-19. SARS-CoV-2 is classified as a biosafety level-3 (BSL-3) agent, impeding the basic research into its biology and the development of effective antivirals. Here, we developed a biosafety level-2 (BSL-2) cell culture system for production of transcription and replication-competent SARS-CoV-2 virus-like-particles (trVLP). This trVLP expresses a reporter gene (GFP) replacing viral nucleocapsid gene (N), which is required for viral genome packaging and virion assembly (SARS-CoV-2 GFP\/\u0394N trVLP). The complete viral life cycle can be achieved and exclusively confined in the cells ectopically expressing SARS-CoV or SARS-CoV-2 N proteins, but not MERS-CoV N. Genetic recombination of N supplied\n                    <jats:italic>in trans<\/jats:italic>\n                    into viral genome was not detected, as evidenced by sequence analysis after one-month serial passages in the N-expressing cells. Moreover, intein-mediated protein trans-splicing approach was utilized to split the viral N gene into two independent vectors, and the ligated viral N protein could function\n                    <jats:italic>in trans<\/jats:italic>\n                    to recapitulate entire viral life cycle, further securing the biosafety of this cell culture model. Based on this BSL-2 SARS-CoV-2 cell culture model, we developed a 96-well format high throughput screening for antivirals discovery. We identified salinomycin, tubeimoside I, monensin sodium, lycorine chloride and nigericin sodium as potent antivirals against SARS-CoV-2 infection. Collectively, we developed a convenient and efficient SARS-CoV-2 reverse genetics tool to dissect the virus life cycle under a BSL-2 condition. This powerful tool should accelerate our understanding of SARS-CoV-2 biology and its antiviral development.\n                  <\/jats:p>","DOI":"10.1371\/journal.ppat.1009439","type":"journal-article","created":{"date-parts":[[2021,3,12]],"date-time":"2021-03-12T13:38:40Z","timestamp":1615556320000},"page":"e1009439","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":195,"title":["A novel cell culture system modeling the SARS-CoV-2 life 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Cliff","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2941-4808","authenticated-orcid":true,"given":"Rong","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiang","family":"Ding","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,3,12]]},"reference":[{"issue":"10223","key":"ppat.1009439.ref001","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1016\/S0140-6736(20)30185-9","article-title":"A novel coronavirus outbreak of global health concern","volume":"395","author":"C Wang","year":"2020","journal-title":"Lancet"},{"issue":"24","key":"ppat.1009439.ref002","doi-asserted-by":"crossref","first-page":"2282","DOI":"10.1056\/NEJMp2009457","article-title":"Drug Evaluation during the Covid-19 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Ianevski","year":"2020","journal-title":"Viruses."},{"issue":"5","key":"ppat.1009439.ref039","doi-asserted-by":"crossref","first-page":"1077","DOI":"10.1016\/j.cell.2020.08.021","article-title":"Emerging Pandemic Diseases: How We Got to COVID-19","volume":"182","author":"DM Morens","year":"2020","journal-title":"Cell"},{"issue":"7816","key":"ppat.1009439.ref040","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1038\/s41586-020-2286-9","article-title":"A SARS-CoV-2 protein interaction map reveals targets for drug repurposing","volume":"583","author":"DE Gordon","year":"2020","journal-title":"Nature"},{"issue":"1","key":"ppat.1009439.ref041","doi-asserted-by":"crossref","first-page":"6041","DOI":"10.1038\/s41467-020-19843-1","article-title":"Nucleocapsid protein of SARS-CoV-2 phase separates into RNA-rich polymerase-containing condensates","volume":"11","author":"A Savastano","journal-title":"Nature Communications. 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Ju","year":"2020","journal-title":"PLoS Pathog."},{"issue":"4","key":"ppat.1009439.ref045","doi-asserted-by":"crossref","first-page":"914","DOI":"10.1016\/j.cell.2020.04.011","article-title":"The Architecture of SARS-CoV-2 Transcriptome","volume":"181","author":"D Kim","year":"2020","journal-title":"Cell."}],"updated-by":[{"DOI":"10.1371\/journal.ppat.1009439","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2021,3,24]],"date-time":"2021-03-24T00:00:00Z","timestamp":1616544000000}}],"container-title":["PLOS 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