An Infectious cDNA Clone of SARS-CoV-2

被引：454

作者：

Xie, Xuping ^{[1
]}

Muruato, Antonio ^{[1
,2
]}

Lokugamage, Kumari G. ^{[2
]}

Narayanan, Krishna ^{[2
]}

Zhang, Xianwen ^{[1
]}

Zou, Jing ^{[1
]}

Liu, Jianying ^{[2
]}

Schindewolf, Craig ^{[2
]}

Bopp, Nathen E. ^{[3
]}

Aguilar, Patricia, V ^{[3
,4
,5
]}

Plante, Kenneth S. ^{[2
,4
]}

Weaver, Scott C. ^{[2
,3
,4
,5
,6
,7
,8
]}

Makino, Shinji ^{[2
,5
,10
]}

LeDuc, James W. ^{[2
,9
]}

Menachery, Vineet D. ^{[2
,3
,5
,7
]}

Shi, Pei-Yong ^{[1
,5
,8
,10
,11
]}

机构：

[1] Univ Texas Med Branch, Dept Biochem & Mol Biol, Galveston, TX 77555 USA

[2] Univ Texas Med Branch, Dept Microbiol & Immunol, Galveston, TX 77555 USA

[3] Univ Texas Med Branch, Dept Pathol, Galveston, TX 77555 USA

[4] Univ Texas Med Branch, World Reference Ctr Emerging Viruses & Arboviruse, Galveston, TX 77555 USA

[5] Univ Texas Med Branch, Inst Human Infect & Immun, Galveston, TX 77555 USA

[6] Univ Texas Med Branch, Inst Translat Sci, Galveston, TX 77555 USA

[7] Univ Texas Med Branch, Ctr Biodef & Emerging Infect Dis, Galveston, TX 77555 USA

[8] Univ Texas Med Branch, Sealy Inst Vaccine Sci, Galveston, TX 77555 USA

[9] Univ Texas Med Branch, Galveston Natl Lab, Galveston, TX 77555 USA

[10] Univ Texas Med Branch, Sealy Ctr Struct Biol & Mol Biophys, Galveston, TX 77555 USA

[11] Univ Texas Med Branch, Dept Pharmacol & Toxicol, Galveston, TX 77555 USA

来源：

CELL HOST & MICROBE | 2020年 / 27卷 / 05期

关键词：

REVERSE GENETICS; SARS;

D O I：

10.1016/j.chom.2020.04.004

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

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 x 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.

引用

页码：841 / +

页数：11

共 22 条

[1]

[Anonymous], VIRUSES

[2]

[Anonymous], BIORXIV

[3]

[Anonymous], 2020, JAMA, DOI DOI 10.1001/JAMA.2020.2648

[4]

[Anonymous], 2020, SARS COV 2 IS SENSIT, DOI DOI 10.1101/2020.03.07.982264

[5] Epidemiological, demographic, and clinical characteristics of 47 cases of Middle East respiratory syndrome coronavirus disease from Saudi Arabia: a descriptive study [J].

Assiri, Abdullah ;

Al-Tawfiq, Jaffar A. ;

Al-Rabeeah, Abdullah A. ;

Al-Rabiah, Fahad A. ;

Al-Hajjar, Sami ;

Al-Barrak, Ali ;

Flemban, Hesham ;

Al-Nassir, Wafa N. ;

Balkhy, Hanan H. ;

Al-Hakeem, Rafat F. ;

Makhdoom, Hatem Q. ;

Zumla, Alimuddin I. ;

Memish, Ziad A. .

LANCET INFECTIOUS DISEASES, 2013, 13 (09) :752-761

[6] The spike glycoprotein of the new coronavirus 2019-nCoV contains a furinlike cleavage site absent in CoV of the same clade [J].

Coutard, B. ;

Valle, C. ;

de lamballerie, X. ;

Canard, B. ;

Seidah, N. G. ;

Decroly, E. .

ANTIVIRAL RESEARCH, 2020, 176

[7] SARS and MERS: recent insights into emerging coronaviruses [J].

de Wit, Emmie ;

van Doremalen, Neeltje ;

Falzarano, Darryl ;

Munster, Vincent J. .

NATURE REVIEWS MICROBIOLOGY, 2016, 14 (08) :523-534

[8] Severe Acute Respiratory Syndrome Coronavirus 2 from Patient with Coronavirus Disease, United States [J].

Harcourt, Jennifer ;

Tamin, Azaibi ;

Lu, Xiaoyan ;

Kamili, Shifaq ;

Sakthivel, Senthil K. ;

Murray, Janna ;

Queen, Krista ;

Tao, Ying ;

Paden, Clinton R. ;

Zhang, Jing ;

Li, Yan ;

Uehara, Anna ;

Wang, Haibin ;

Goldsmith, Cynthia ;

Bullock, Hannah A. ;

Wang, Lijuan ;

Whitaker, Brett ;

Lynch, Brian ;

Gautam, Rashi ;

Schindewolf, Craig ;

Lokugamage, Kumari G. ;

Scharton, Dionna ;

Plante, Jessica A. ;

Mirchandani, Divya ;

Widen, Steven G. ;

Narayanan, Krishna ;

Makino, Shinji ;

Ksiazek, Thomas G. ;

Plante, Kenneth S. ;

Weaver, Scott C. ;

Lindstrom, Stephen ;

Tong, Suxiang ;

Menachery, Vineet D. ;

Thornburg, Natalie J. .

EMERGING INFECTIOUS DISEASES, 2020, 26 (06) :1266-1273

[9] First Case of 2019 Novel Coronavirus in the United States [J].

Holshue, Michelle L. ;

DeBolt, Chas ;

Lindquist, Scott ;

Lofy, Kathy H. ;

Wiesman, John ;

Bruce, Hollianne ;

Spitters, Christopher ;

Ericson, Keith ;

Wilkerson, Sara ;

Tural, Ahmet ;

Diaz, George ;

Cohn, Amanda ;

Fox, LeAnne ;

Patel, Anita ;

Gerber, Susan I. ;

Kim, Lindsay ;

Tong, Suxiang ;

Lu, Xiaoyan ;

Lindstrom, Steve ;

Pallansch, Mark A. ;

Weldon, William C. ;

Biggs, Holly M. ;

Uyeki, Timothy M. ;

Pillai, Satish K. .

NEW ENGLAND JOURNAL OF MEDICINE, 2020, 382 (10) :929-936

[10] SARS-like WIV1-CoV poised for human emergence [J].

Menachery, Vineet D. ;

Yount, Boyd L., Jr. ;

Sims, Amy C. ;

Debbink, Kari ;

Agnihothram, Sudhakar S. ;

Gralinski, Lisa E. ;

Graham, Rachel L. ;

Scobey, Trevor ;

Plante, Jessica A. ;

Royal, Scott R. ;

Swanstrom, Jesica ;

Sheahan, Timothy P. ;

Pickles, Raymond J. ;

Corti, Davide ;

Randell, Scott H. ;

Lanzavecchia, Antonio ;

Marasco, Wayne A. ;

Baric, Ralph S. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2016, 113 (11) :3048-3053

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