Journal
CELL HOST & MICROBE
Volume 27, Issue 5, Pages 841-+Publisher
CELL PRESS
DOI: 10.1016/j.chom.2020.04.004
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Funding
- NIA
- NIAID of the NIH [U19AI100625, R00AG049092, R24AI120942, AI114657, AI146081, 5UC7AI094660]
- STARs Award by the University of Texas System
- McLaughlin Fellowship Fund at UTMB
- IHII Pilot grant
- NIH [AI142759, AI145617, AI127744, AI136126, AI134907, UL1TR001439]
- Kleberg Foundation
- John S. Dunn Foundation
- Amon G. Carter Foundation
- Gilson Longenbaugh Foundation
- Summerfield Robert Foundation
- Clinical and Translational Science Award NRSA (TL1) Training Core from the NIH [TL1TR001440]
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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 x 10(6) 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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