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

doi:10.1016/j.chom.2020.04.004

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 journal › Article › peer-review

427 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 language	English (US)
Pages (from-to)	841-848.e3
Journal	Cell Host and Microbe
Volume	27
Issue number	5
DOIs	https://doi.org/10.1016/j.chom.2020.04.004
State	Published - May 13 2020

Keywords

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

ASJC Scopus subject areas

Parasitology
Microbiology
Virology

Access to Document

10.1016/j.chom.2020.04.004

Cite this

@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 = "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",

}

TY - JOUR

T1 - An Infectious cDNA Clone of SARS-CoV-2

AU - Xie, Xuping

AU - Muruato, Antonio

AU - Lokugamage, Kumari G.

AU - Narayanan, Krishna

AU - Zhang, Xianwen

AU - Zou, Jing

AU - Liu, Jianying

AU - Schindewolf, Craig

AU - Bopp, Nathen E.

AU - Aguilar, Patricia V.

AU - Plante, Kenneth S.

AU - Weaver, Scott C.

AU - Makino, Shinji

AU - LeDuc, James W.

AU - Menachery, Vineet D.

AU - Shi, Pei Yong

PY - 2020/5/13

Y1 - 2020/5/13

N2 - 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.

AB - 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.

KW - COVID-19

KW - SARS-CoV

KW - SARS-CoV-2

KW - antiviral

KW - coronavirus

KW - vaccine

UR - http://www.scopus.com/inward/record.url?scp=85083529350&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85083529350&partnerID=8YFLogxK

U2 - 10.1016/j.chom.2020.04.004

DO - 10.1016/j.chom.2020.04.004

M3 - Article

C2 - 32289263

AN - SCOPUS:85083529350

SN - 1931-3128

VL - 27

SP - 841-848.e3

JO - Cell Host and Microbe

JF - Cell Host and Microbe

IS - 5

ER -

An Infectious cDNA Clone of SARS-CoV-2

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this