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An Infectious cDNA Clone of SARS-CoV-2
Open accessAbstract
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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“Together with recent reports of the preserved neutralization of SARS-CoV-2 with key S mutations found in the B.1.1.7 and the B.1.351lineage 39,40 , and neutralization of pseudovirus bearing the full set of mutations of B.1.1.7 S 41 , our data indicate broad BNT162b2-elicited immune recognition 2,13,14 .”
From BNT162b2 vaccine induces neutralizing antibodies and poly-specific T cells in humans (2021) · cites this paper as [40]
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“Two variants had either eight (D80A/D215G/Δ242-244/ K417N/E484K/N501Y/D614G/A701V) or ten mutations (L18F/D80A/ D215G/R246I/Δ242-244/K417N/E484K/N501Y/D614G/A701V) found in S of the B.1.351lineage (also referred to as 20C/501Y.V2) 40 .”
From BNT162b2 vaccine induces neutralizing antibodies and poly-specific T cells in humans (2021) · cites this paper as [40]
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“This mutation is located in the viral receptor-binding domain (RBD) for cell entry, increases binding to the angiotensin-converting enzyme 2 receptor”
From Neutralization of SARS-CoV-2 spike 69/70 deletion, E484K and N501Y variants by BNT162b2 vaccine-elicited sera (2021) · cites this paper as [5]
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“and enables the virus to expand its host range to infect mice 5,6 .”
From Neutralization of SARS-CoV-2 spike 69/70 deletion, E484K and N501Y variants by BNT162b2 vaccine-elicited sera (2021) · cites this paper as [5]
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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
- ORCID
- Venue
- Cell Host & Microbe
- Publisher
- Elsevier BV
- Published
- 2020-05
- Volume
- 27
- Issue
- 5
- Pages
- 841-848.e3
- ISSN
- 1931-3128 (ISSN-L)
- PMC
- PMC7153529
- 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
- bronze
- doi.org
- Licence
- 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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