Your browser doesn't support javascript.
An Infectious cDNA Clone of SARS-CoV-2.
Xie, Xuping; Muruato, Antonio; Lokugamage, Kumari G; Narayanan, Krishna; Zhang, Xianwen; Zou, Jing; Liu, Jianying; Schindewolf, Craig; Bopp, Nathen E; Aguilar, Patricia V; Plante, Kenneth S; Weaver, Scott C; Makino, Shinji; LeDuc, James W; Menachery, Vineet D; Shi, Pei-Yong.
  • Xie X; Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, USA. Electronic address: xuxie@UTMB.edu.
  • Muruato A; Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, USA; Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA.
  • Lokugamage KG; Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA.
  • Narayanan K; Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA.
  • Zhang X; Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, USA.
  • Zou J; Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, USA.
  • Liu J; Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA.
  • Schindewolf C; Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA.
  • Bopp NE; Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA.
  • Aguilar PV; Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA; World Reference Center for Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infection and Immunity, University of Texas Medical Branch, Galveston, TX, USA.
  • Plante KS; Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; World Reference Center for Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, TX, USA.
  • Weaver SC; Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; World Reference Center for Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infection and Immunity, University of Texas Medical Branch, Galv
  • Makino S; Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infection and Immunity, University of Texas Medical Branch, Galveston, TX, USA; Sealy Center for Structural Biology & Molecular Biophysics, University of Texas Medical Branch, G
  • LeDuc JW; Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; Galveston National Laboratory, University of Texas Medical Branch, Galveston, TX, USA.
  • Menachery VD; Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infection and Immunity, University of Texas Medical Branch, Galveston, TX, USA; Department of Pathology and Center for Biodefense & Emerging Infectious Diseases, University of T
  • Shi PY; Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infection and Immunity, University of Texas Medical Branch, Galveston, TX, USA; Sealy Institute for Vaccine Sciences, University of Texas Medical Branch, Galveston, TX, USA;
Cell Host Microbe ; 27(5): 841-848.e3, 2020 05 13.
Article in English | MEDLINE | ID: covidwho-716611
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.
Subject(s)
Keywords
Fulltext
Print
XML
PubMed Links
Search on Google

Full text: Available Collection: International databases Database: MEDLINE Main subject: Pneumonia, Viral / DNA, Complementary / Coronavirus Infections / Organisms, Genetically Modified / Betacoronavirus Type of study: Experimental Studies / Prognostic study Limits: Animals Language: English Journal: Cell Host Microbe Journal subject: Microbiology Year: 2020 Document Type: Article

Similar

MEDLINE
LILACS
LIS
Fulltext
Print
XML
PubMed Links
Search on Google

Full text: Available Collection: International databases Database: MEDLINE Main subject: Pneumonia, Viral / DNA, Complementary / Coronavirus Infections / Organisms, Genetically Modified / Betacoronavirus Type of study: Experimental Studies / Prognostic study Limits: Animals Language: English Journal: Cell Host Microbe Journal subject: Microbiology Year: 2020 Document Type: Article