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Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome
mBio ( IF 5.1 ) Pub Date : 2020-09-25 , DOI: 10.1128/mbio.02168-20 Chengjin Ye 1 , Kevin Chiem 1 , Jun-Gyu Park 1 , Fatai Oladunni 1, 2 , Roy Nelson Platt 1 , Tim Anderson 1 , Fernando Almazan 3 , Juan Carlos de la Torre 4 , Luis Martinez-Sobrido 1
mBio ( IF 5.1 ) Pub Date : 2020-09-25 , DOI: 10.1128/mbio.02168-20 Chengjin Ye 1 , Kevin Chiem 1 , Jun-Gyu Park 1 , Fatai Oladunni 1, 2 , Roy Nelson Platt 1 , Tim Anderson 1 , Fernando Almazan 3 , Juan Carlos de la Torre 4 , Luis Martinez-Sobrido 1
Affiliation
Infectious coronavirus (CoV) disease 2019 (COVID-19) emerged in the city of Wuhan (China) in December 2019, causing a pandemic that has dramatically impacted public health and socioeconomic activities worldwide. A previously unknown coronavirus, severe acute respiratory syndrome CoV-2 (SARS-CoV-2), has been identified as the causative agent of COVID-19. To date, there are no U.S. Food and Drug Administration (FDA)-approved vaccines or therapeutics available for the prevention or treatment of SARS-CoV-2 infection and/or associated COVID-19 disease, which has triggered a large influx of scientific efforts to develop countermeasures to control SARS-CoV-2 spread. To contribute to these efforts, we have developed an infectious cDNA clone of the SARS-CoV-2 USA-WA1/2020 strain based on the use of a bacterial artificial chromosome (BAC). Recombinant SARS-CoV-2 (rSARS-CoV-2) was readily rescued by transfection of the BAC into Vero E6 cells. Importantly, BAC-derived rSARS-CoV-2 exhibited growth properties and plaque sizes in cultured cells comparable to those of the natural SARS-CoV-2 isolate. Likewise, rSARS-CoV-2 showed levels of replication similar to those of the natural isolate in nasal turbinates and lungs of infected golden Syrian hamsters. This is, to our knowledge, the first BAC-based reverse genetics system for the generation of infectious rSARS-CoV-2 that displays features in vivo similar to those of a natural viral isolate. This SARS-CoV-2 BAC-based reverse genetics will facilitate studies addressing several important questions in the biology of SARS-CoV-2, as well as the identification of antivirals and development of vaccines for the treatment of SARS-CoV-2 infection and associated COVID-19 disease.
更新日期：2020-10-28
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