Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome

ABSTRACT 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...

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Autores principales:	Chengjin Ye, Kevin Chiem, Jun-Gyu Park, Fatai Oladunni, Roy Nelson Platt, Tim Anderson, Fernando Almazan, Juan Carlos de la Torre, Luis Martinez-Sobrido
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	BAC COVID-19 SARS-CoV-2 coronavirus hamsters recombinant virus Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/b595b2ca0ebe45ea9b2ab483468d4e53
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spelling	oai:doaj.org-article:b595b2ca0ebe45ea9b2ab483468d4e532021-11-15T16:19:07ZRescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome10.1128/mBio.02168-202150-7511https://doaj.org/article/b595b2ca0ebe45ea9b2ab483468d4e532020-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02168-20https://doaj.org/toc/2150-7511ABSTRACT 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. IMPORTANCE The pandemic coronavirus (CoV) disease 2019 (COVID-19) caused by severe acute respiratory syndrome CoV-2 (SARS-CoV-2) is a major threat to global human health. To date, there are no approved prophylactics or therapeutics available for COVID-19. Reverse genetics is a powerful approach to understand factors involved in viral pathogenesis, antiviral screening, and vaccine development. In this study, we describe the feasibility of generating recombinant SARS-CoV-2 (rSARS-CoV-2) by transfection of a single bacterial artificial chromosome (BAC). Importantly, rSARS-CoV-2 possesses the same phenotype as the natural isolate in vitro and in vivo. This is the first description of a BAC-based reverse genetics system for SARS-CoV-2 and the first time that an rSARS-CoV-2 isolate has been shown to be phenotypically identical to a natural isolate in a validated animal model of SARS-CoV-2 infection. The BAC-based reverse genetics approach will facilitate the study of SARS-CoV-2 and the development of prophylactics and therapeutics for the treatment of COVID-19.Chengjin YeKevin ChiemJun-Gyu ParkFatai OladunniRoy Nelson PlattTim AndersonFernando AlmazanJuan Carlos de la TorreLuis Martinez-SobridoAmerican Society for MicrobiologyarticleBACCOVID-19SARS-CoV-2coronavirushamstersrecombinant virusMicrobiologyQR1-502ENmBio, Vol 11, Iss 5 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	BAC COVID-19 SARS-CoV-2 coronavirus hamsters recombinant virus Microbiology QR1-502
spellingShingle	BAC COVID-19 SARS-CoV-2 coronavirus hamsters recombinant virus Microbiology QR1-502 Chengjin Ye Kevin Chiem Jun-Gyu Park Fatai Oladunni Roy Nelson Platt Tim Anderson Fernando Almazan Juan Carlos de la Torre Luis Martinez-Sobrido Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome
description	ABSTRACT 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. IMPORTANCE The pandemic coronavirus (CoV) disease 2019 (COVID-19) caused by severe acute respiratory syndrome CoV-2 (SARS-CoV-2) is a major threat to global human health. To date, there are no approved prophylactics or therapeutics available for COVID-19. Reverse genetics is a powerful approach to understand factors involved in viral pathogenesis, antiviral screening, and vaccine development. In this study, we describe the feasibility of generating recombinant SARS-CoV-2 (rSARS-CoV-2) by transfection of a single bacterial artificial chromosome (BAC). Importantly, rSARS-CoV-2 possesses the same phenotype as the natural isolate in vitro and in vivo. This is the first description of a BAC-based reverse genetics system for SARS-CoV-2 and the first time that an rSARS-CoV-2 isolate has been shown to be phenotypically identical to a natural isolate in a validated animal model of SARS-CoV-2 infection. The BAC-based reverse genetics approach will facilitate the study of SARS-CoV-2 and the development of prophylactics and therapeutics for the treatment of COVID-19.
format	article
author	Chengjin Ye Kevin Chiem Jun-Gyu Park Fatai Oladunni Roy Nelson Platt Tim Anderson Fernando Almazan Juan Carlos de la Torre Luis Martinez-Sobrido
author_facet	Chengjin Ye Kevin Chiem Jun-Gyu Park Fatai Oladunni Roy Nelson Platt Tim Anderson Fernando Almazan Juan Carlos de la Torre Luis Martinez-Sobrido
author_sort	Chengjin Ye
title	Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome
title_short	Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome
title_full	Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome
title_fullStr	Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome
title_full_unstemmed	Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome
title_sort	rescue of sars-cov-2 from a single bacterial artificial chromosome
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/b595b2ca0ebe45ea9b2ab483468d4e53
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Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome

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