[PDF][PDF] An infectious cDNA clone of SARS-CoV-2

X Xie, A Muruato, KG Lokugamage, K Narayanan… - Cell host & …, 2020 - cell.com
X Xie, A Muruato, KG Lokugamage, K Narayanan, X Zhang, J Zou, J Liu, C Schindewolf
Cell host & microbe, 2020cell.com
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× 10 6 plaque …
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.
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