An Infectious cDNA Clone of SARS-CoV-2

被引：433

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; CORONAVIRUS; SARS; EXPRESSION;

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 50 条

[41] Genetics and genomics of SARS-CoV-2: A review of the literature with the special focus on genetic diversity and SARS-CoV-2 genome detection
Rahimi, Azadeh
Mirzazadeh, Azin
Tavakolpour, Soheil
GENOMICS, 2021, 113 (01) : 1221 - 1232
[42] There is no "origin" to SARS-CoV-2
Frutos, Roger
Pliez, Olivier
Gavotte, Laurent
Devaux, Christian A.
ENVIRONMENTAL RESEARCH, 2022, 207
[43] Investigation of SARS-CoV-2 IgG Binding Capability to Variants of the SARS-CoV-2 Virus
Johnson, Lucy
De Gascun, Cillian F.
Hassan, Jaythoon
VIRAL IMMUNOLOGY, 2024, 37 (08) : 404 - 410
[44] Should we expect neurological symptoms in the SARS-CoV-2 epidemic?
Matias-Guiu, J.
Gomez-Pinedo, U.
Montero-Escribano, P.
Gomez-Iglesias, P.
Porta-Etessam, J.
Matias-Guiu, J. A.
NEUROLOGIA, 2020, 35 (03): : 170 - 175
[45] Immune Responses in SARS-CoV-2, SARS-CoV, and MERS-CoV Infections: A Comparative Review
Irani, Soussan
INTERNATIONAL JOURNAL OF PREVENTIVE MEDICINE, 2022, 13 (01) : 45
[46] SARS-CoV-2 infection protects against rechallenge in rhesus macaques
Chandrashekar, Abishek
Liu, Jinyan
Martinot, Amanda J.
McMahan, Katherine
Mercado, Noe B.
Peter, Lauren
Tostanoski, Lisa H.
Yu, Jingyou
Maliga, Zoltan
Nekorchuk, Michael
Busman-Sahay, Kathleen
Terry, Margaret
Wrijil, Linda M.
Ducat, Sarah
Martinez, David R.
Atyeo, Caroline
Fischinger, Stephanie
Burke, John S.
Slein, Matthew D.
Pessaint, Laurent
Van Ry, Alex
Greenhouse, Jack
Taylor, Tammy
Blade, Kelvin
Cook, Anthony
Finneyfrock, Brad
Brown, Renita
Teow, Elyse
Velasco, Jason
Zahn, Roland
Wegmann, Frank
Abbink, Peter
Bondzie, Esther A.
Dagotto, Gabriel
Gebre, Makda S.
He, Xuan
Jacob-Dolan, Catherine
Kordana, Nicole
Li, Zhenfeng
Lifton, Michelle A.
Mahrokhian, Shant H.
Maxfield, Lori F.
Nityanandam, Ramya
Nkolola, Joseph P.
Schmidt, Aaron G.
Miller, Andrew D.
Baric, Ralph S.
Alter, Galit
Sorger, Peter K.
Estes, Jacob D.
SCIENCE, 2020, 369 (6505) : 812 - +
[47] Determination of the cycle threshold value of the Xpert Xpress SARS-CoV-2/Flu/RSV test that corresponds to the presence of infectious SARS-CoV-2 in anterior nasal swabs
Relich, Ryan F.
Van Benten, Kayla
Lei, Guang-Sheng
Robinson, Christopher M.
Carozza, Mariel
Sahoo, Malaya K.
Huang, Chunhong
Solis, Daniel
Sibai, Mamdouh
Myers, Christopher A.
Sikorski, Cynthia
Balagot, Caroline
Yang, David
Pinsky, Benjamin A.
Loeffelholz, Michael J.
MICROBIOLOGY SPECTRUM, 2024, 12 (04):
[48] Mapping the technological landscape of SARS, MERS, and SARS-CoV-2 vaccines
Nascimento Junior, Jose Adao Carvalho
Santos, Anamaria Mendonca
Cavalcante, Rafael Ciro Marques
Quintans-Junior, Lucindo Jose
Walker, Cristiani Isabel Bandero
Borges, Lysandro Pinto
Frank, Luiza Abrahao
Serafini, Mairim Russo
DRUG DEVELOPMENT AND INDUSTRIAL PHARMACY, 2021, 47 (04) : 673 - 684
[49] Comparing SARS-CoV-2 with SARS-CoV and influenza pandemics
Petersen, Eskild
Koopmans, Marion
Go, Unyeong
Hamer, Davidson H.
Petrosillo, Nicola
Castelli, Francesco
Storgaard, Merete
Al Khalili, Sulien
Simonsen, Lone
LANCET INFECTIOUS DISEASES, 2020, 20 (09) : E238 - E244
[50] Construction of Non-infectious SARS-CoV-2 Replicons and Their Application in Drug Evaluation
Wang, Bei
Zhang, Chongyang
Lei, Xiaobo
Ren, Lili
Huang, He
Wang, Jianwei
Zhao, Zhendong
VIROLOGICA SINICA, 2021, 36 (05) : 890 - 900

← 1 2 3 4 5 →