An Infectious cDNA Clone of SARS-CoV-2

By Xuping Xie, Antonio E. Muruato, K. Lokugamage, Krishna R. Narayanan, Xianwen Zhang, Jing Zou, Jianying Liu, Craig Schindewolf, Nathen E. Bopp, P. Aguilar and 6 more. Cell Host and Microbe, 2020

644 citations recorded by the source corpus.

Summary 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.

Subfields: Animal Science and Zoology, Infectious Diseases

View Paper: https://www.semanticscholar.org/paper/5a591cae690b53a9ca95cc21e7b7f35ac8a3d049

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In Animal Science and Zoology

1. F. Krammer

In Infectious Diseases

1. Alessandro Sette
2. Shane Crotty
3. F. Krammer

Citations in the corpus

• Evasion of Type I Interferon by SARS-CoV-2 (2020, methodology): “Each gene was derived from PCR using infectious cDNA clone plasmids (Xie et al., 2020a), and fused with a Flag-tag at C-terminal.”
• Targets of T Cell Responses to SARS-CoV-2 Coronavirus in Humans with COVID-19 Disease and Unexposed Individuals (2020, positive): “Accounting for the relative abundance of subgenomic RNAs (Figure 6A) (Irigoyen et al., 2016; Snijder et al., 2003; Xie et al., 2020), the ORFs were re-ordered based on predicted protein abundance (Figure 6B).”