An Infectious cDNA Clone of SARS-CoV-2

• Published in: Cell host & microbe Vol. 27; no. 5; pp. 841 - 848.e3
• Main Authors: Xie, Xuping, Muruato, Antonio, Lokugamage, Kumari G., Narayanan, Krishna, Zhang, Xianwen, Zou, Jing, Liu, Jianying, Schindewolf, Craig, Bopp, Nathen E., Aguilar, Patricia V., Plante, Kenneth S., Weaver, Scott C., Makino, Shinji, LeDuc, James W., Menachery, Vineet D., Shi, Pei-Yong
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 13.05.2020
• Subjects: Animals; antiviral; Antiviral Agents - therapeutic use; Betacoronavirus - genetics; Betacoronavirus - pathogenicity; Chlorocebus aethiops; Clone Cells; coronavirus; Coronavirus Infections - drug therapy; Coronavirus Infections - virology; COVID-19; DNA, Complementary - genetics; Genes, Reporter - genetics; Genome, Viral - genetics; Interferons - therapeutic use; Organisms, Genetically Modified - genetics; Organisms, Genetically Modified - pathogenicity; Pandemics; Pneumonia, Viral - drug therapy; Pneumonia, Viral - virology; RNA, Viral - genetics; SARS-CoV; SARS-CoV-2; vaccine; Vero Cells - virology; Virus Replication - physiology; COVID-19; coronavirus; SARS-CoV-2; vaccine; SARS-CoV; antiviral
• ISSN: 1931-3128; 1934-6069; 1934-6069
• DOI: 10.1016/j.chom.2020.04.004

The ongoing pandemic of COVID-19, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), underscores the urgency to develop experimental systems for studying this virus and identifying countermeasures. We report a reverse genetic system for SARS-CoV-2. Seven complimentary DNA (cDNA) fragments spanning the SARS-CoV-2 genome were assembled into a full-genome cDNA. RNA transcribed from the full-genome cDNA was highly infectious after electroporation into cells, producing 2.9 × 106 plaque-forming unit (PFU)/mL of virus. Compared with a clinical isolate, the infectious-clone-derived SARS-CoV-2 (icSARS-CoV-2) exhibited similar plaque morphology, viral RNA profile, and replication kinetics. Additionally, icSARS-CoV-2 retained engineered molecular markers and did not acquire other mutations. We generated a stable mNeonGreen SARS-CoV-2 (icSARS-CoV-2-mNG) by introducing this reporter gene into ORF7 of the viral genome. icSARS-CoV-2-mNG was successfully used to evaluate the antiviral activities of interferon (IFN). Collectively, the reverse genetic system and reporter virus provide key reagents to study SARS-CoV-2 and develop countermeasures. [Display omitted] •A reverse genetic system has been established for SARS-CoV-2•Recombinant SARS-CoV-2 replicates as efficiently as the original clinical isolate•A stable mNeonGreen reporter SARS-CoV-2 has been developed•The mNeonGreen SARS-CoV-2 can be used to screen antiviral inhibitors Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a devastating global pandemic. Xie et al. generated an infectious cDNA clone of SARS-CoV-2 and a mNeonGreen reporter virus. Recombinant SARS-CoV-2 and reporter virus replicate as efficiently as the original clinical isolate.