@article{c8ba8fbe8b4d463e85c932fe5f6f877e,
title = "An Infectious cDNA Clone of SARS-CoV-2",
abstract = "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.",
keywords = "COVID-19, SARS-CoV, SARS-CoV-2, antiviral, coronavirus, vaccine",
author = "Xuping Xie and Antonio Muruato and Lokugamage, {Kumari G.} and Krishna Narayanan and Xianwen Zhang and Jing Zou and Jianying Liu and Craig Schindewolf and Bopp, {Nathen E.} and Aguilar, {Patricia V.} and Plante, {Kenneth S.} and Weaver, {Scott C.} and Shinji Makino and LeDuc, {James W.} and Menachery, {Vineet D.} and Shi, {Pei Yong}",
note = "Funding Information: We thank Natalie Thornburg and other colleagues from the Centers for Disease Control and Prevention for providing the clinical virus isolate. We also thank colleagues at University of Texas Medical Branch (UTMB) for support and discussions. Research was supported by grants from NIA and NIAID of the NIH (U19AI100625 and R00AG049092 to V.D.M.; R24AI120942 (WRCEVA) to S.C.W.; AI114657 and AI146081 to S.M.; and 5UC7AI094660 to J.W.L.). Research was also supported by a STARs Award provided by the University of Texas System to V.D.M. trainee funding provided by the McLaughlin Fellowship Fund at UTMB, and an IHII Pilot grant to S.M. P.-Y.S. was supported by NIH grants AI142759, AI145617, AI127744, AI136126, AI134907 and UL1TR001439 and by awards from the Kleberg Foundation, the John S. Dunn Foundation, the Amon G. Carter Foundation, the Gilson Longenbaugh Foundation, and the Summerfield Robert Foundation. A.M. is supported by a Clinical and Translational Science Award NRSA (TL1) Training Core (TL1TR001440) from the NIH. Conceptualization, X.X. V.D.M. and P.-Y.S.; Methodology, X.X. A.M. K.G.L. K.N. X.Z. J.Z. J.L. C.S. N.B. P.A. K.S.P. S.W, S.M. J.W.L. V.D.M, and P.-Y.S.; Investigation, X.X. A.M. K.G.L. K.N. X.Z. J.Z. J.L. C.S. N.B. and P.A.; Resources, K.S.P. S.W. and C.-T.K.T.; Data Curation, X.X. A.M. K.G.L. K.N. J.L. and N.B.; Writing-Original Draft, X.X. K.N. V.D.M. and P.-Y.S.; Writing-Review & Editing, X.X. V.D.M. and P.-Y.S.; Visualization, X.X. A.M. K.G.L. N.B. and P.-Y.S.; Supervision, X.X. V.D.M. and P.-Y.S.; Funding Acquisition, P.A. S.W. S.J. J.W.L. V.D.M. and P.-Y.S. X.X. V.D.M, and P.-Y.S. have filed a provisional patent on the reverse genetic system of SARS-CoV-2. Other authors have no conflicts of interest to declare. Funding Information: We thank Natalie Thornburg and other colleagues from the Centers for Disease Control and Prevention for providing the clinical virus isolate. We also thank colleagues at University of Texas Medical Branch (UTMB) for support and discussions. Research was supported by grants from NIA and NIAID of the NIH ( U19AI100625 and R00AG049092 to V.D.M.; R24AI120942 (WRCEVA) to S.C.W.; AI114657 and AI146081 to S.M.; and 5UC7AI094660 to J.W.L.). Research was also supported by a STARs Award provided by the University of Texas System to V.D.M., trainee funding provided by the McLaughlin Fellowship Fund at UTMB, and an IHII Pilot grant to S.M. P.-Y.S. was supported by NIH grants AI142759 , AI145617 , AI127744 , AI136126 , AI134907 and UL1TR001439 and by awards from the Kleberg Foundation , the John S. Dunn Foundation , the Amon G. Carter Foundation , the Gilson Longenbaugh Foundation , and the Summerfield Robert Foundation . A.M. is supported by a Clinical and Translational Science Award NRSA (TL1) Training Core ( TL1TR001440 ) from the NIH. Publisher Copyright: {\textcopyright} 2020 Elsevier Inc.",
year = "2020",
month = may,
day = "13",
doi = "10.1016/j.chom.2020.04.004",
language = "English (US)",
volume = "27",
pages = "841--848.e3",
journal = "Cell Host and Microbe",
issn = "1931-3128",
publisher = "Cell Press",
number = "5",
}