An Infectious cDNA Clone of SARS-CoV-2

Xuping Xie, Antonio Muruato, Kumari G. Lokugamage, Krishna Narayanan, Xianwen Zhang, Jing Zou, Jianying Liu, Craig Schindewolf, Nathen E. Bopp, Patricia V. Aguilar, Kenneth S. Plante, Scott C. Weaver, Shinji Makino, James W. LeDuc, Vineet D. Menachery, Pei Yong Shi

Research output: Contribution to journalArticle

3 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)841-848.e3
JournalCell Host and Microbe
Volume27
Issue number5
DOIs
StatePublished - May 13 2020

Keywords

  • COVID-19
  • SARS-CoV
  • SARS-CoV-2
  • antiviral
  • coronavirus
  • vaccine

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Virology

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  • Cite this

    Xie, X., Muruato, A., Lokugamage, K. G., Narayanan, K., Zhang, X., Zou, J., Liu, J., Schindewolf, C., Bopp, N. E., Aguilar, P. V., Plante, K. S., Weaver, S. C., Makino, S., LeDuc, J. W., Menachery, V. D., & Shi, P. Y. (2020). An Infectious cDNA Clone of SARS-CoV-2. Cell Host and Microbe, 27(5), 841-848.e3. https://doi.org/10.1016/j.chom.2020.04.004