Cited by 649SARS-CoV-2 and COVID-19 ResearchAnimal Virus Infections StudiesSARS-CoV-2 detection and testing

The DOI matches its Crossref record; 28 of the 30 references it cites were found in a registry or an index.

An Infectious cDNA Clone of SARS-CoV-2

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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 × 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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Crossref · 10.1016/j.chom.2020.04.004

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  1. Lokugamage, 2020, SARS-CoV-2 sensitive to type I interferon pretreatment, bioRxiv

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Metadata

Authors
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.
Affiliations
The University of Texas Medical Branch at Galveston (US)
Corresponding author
  • Xie, Xuping
  • Menachery, Vineet D.
  • Shi, Pei-Yong
Venue
Cell Host & Microbe
Publisher
Elsevier BV
Published
2020-05
Volume
27
Issue
5
Pages
841-848.e3
ISSN
1931-3128 (ISSN-L)
OpenAlex
W3015751519
Cited by
647
References
30
Citations by year
2026: 30 · 2025: 47 · 2024: 54 · 2023: 94 · 2022: 162 · 2021: 180 · 2020: 80
FWCI
16.94
Type
journal article
Language
en
Open access
Locations
  • Cell Host & Microbe (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Centers for Disease Control and Prevention
  • National Institute on Aging
  • National Institute of Allergy and Infectious Diseases
  • Amon G. Carter Foundation
  • Summerfield G. Roberts Foundation
  • National Institutes of Health (5UC7AI094660, AI114657, AI146081, R00AG049092, R24AI120942, U19AI100625)
  • University of Texas Medical Branch
  • Gillson Longenbaugh Foundation
  • Robert J. Kleberg, Jr. and Helen C. Kleberg Foundation
  • John S. Dunn Foundation
  • University of Texas System (AI127744, AI134907, AI136126, AI142759, AI145617, UL1TR001439)
Topics
  • Health Sciences › Medicine › Infectious Diseases › SARS-CoV-2 and COVID-19 Research
  • Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Animal Virus Infections Studies
  • Health Sciences › Medicine › Infectious Diseases › SARS-CoV-2 detection and testing
Keywords
SARS-CoV-2, infectious clone, reverse genetics, reporter virus, ORF7, interferon antiviral activity, viral replication kinetics, plaque morphology, COVID-19 pandemic
MeSH terms
Betacoronavirus, COVID-19, SARS-CoV-2, Animals, Antiviral Agents, Chlorocebus aethiops, Clone Cells, Interferons, Pneumonia, Viral, RNA, Viral, Vero Cells, Virus Replication, Genome, Viral, Genes, Reporter, DNA, Complementary, Coronavirus Infections, Organisms, Genetically Modified, Pandemics

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