An Infectious cDNA Clone of SARS-CoV-2

Cell Host & Microbe
Xuping XiePei-Yong Shi

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.

Citations

Jun 25, 2020·The Journal of General Virology·Severino Jefferson Ribeiro da SilvaLindomar Pena
Jul 30, 2020·PLoS Pathogens·Margarida Sa RiberoSébastien Nisole
Jul 24, 2020·Journal of Virology·Abigail VanderheidenMehul S Suthar
Aug 14, 2020·Nature·Mark J MulliganKathrin U Jansen
Sep 27, 2020·MBio·Chengjin YeLuis Martinez-Sobrido
Sep 30, 2020·The New England Journal of Medicine·Evan J AndersonUNKNOWN mRNA-1273 Study Group
Jul 17, 2020·Nature Nanotechnology·Matthew D ShinNicole F Steinmetz
Aug 15, 2020·Nature Communications·Antonio E MuruatoPei-Yong Shi
Jul 2, 2020·International Journal of Molecular Sciences·Alejandro LlanesRicardo Lleonart
Sep 25, 2020·Journal of Translational Medicine·Francesca CaccuriArnaldo Caruso
Sep 9, 2020·PLoS Biology·Nicole A P LiebermanAlexander L Greninger
Oct 14, 2020·Current Protocols in Microbiology·Kevin ChiemLuis Martinez-Sobrido
Oct 31, 2020·Wiener klinische Wochenschrift·Franz X Heinz, Karin Stiasny
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Nov 6, 2020·PloS One·Eric C RouchkaDonghoon Chung
Nov 21, 2020·Current Protocols in Immunology·Abigail VanderheidenMehul S Suthar
Nov 27, 2020·Proceedings of the National Academy of Sciences of the United States of America·Ruochen ZangSiyuan Ding
Oct 28, 2020·Nature·Jessica A PlantePei-Yong Shi
Dec 19, 2020·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·Hongjie Xia, Pei-Yong Shi
Dec 29, 2020·Advanced Drug Delivery Reviews·Jee Young ChungYoung Jik Kwon
Nov 2, 2020·Diagnostic Microbiology and Infectious Disease·Xuping XiePing Ren
Dec 15, 2020·Journal of Molecular Biology·Takamitsu HattoriShohei Koide
Sep 28, 2020·Cell Reports·Hongjie XiaPei-Yong Shi
Jan 28, 2021·Scientific Reports·Tomohiro KotakiMasanori Kameoka
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Feb 13, 2021·AIMS Public Health·Srikanth UmakanthanMaryann Bukelo
Jan 7, 2021·Nature Structural & Molecular Biology·Robert J EdwardsPriyamvada Acharya

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Methods Mentioned

BETA
in vitro transcription
transfection
Fluorescence microscopy
PCR

Software Mentioned

ImageJ
GENEWIZ
Prism
GraphPad Prism
GraphPad
Adobe illustrator

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