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Development of a Single-Cycle Infectious SARS-CoV-2 Virus Replicon Particle System for Use in Biosafety Level 2 Laboratories

Johnny Malicoat, Senthamizharasi Manivasagam, Sonia Zuñiga, Isabel Sola, Dianne McCabe, Lijun Rong, Stanley Perlman, Luis Enjuanes, Balaji Manicassamy

2021Journal of Virology40 citationsDOIOpen Access PDF

Abstract

Due to the highly contagious nature of SARS-CoV-2 and the lack of immunity in the human population, research on SARS-CoV-2 has been restricted to biosafety level 3 laboratories. This has greatly limited participation of the broader scientific community in SARS-CoV-2 research and thus has hindered the development of vaccines and antiviral drugs. By deleting the essential spike gene in the viral genome, we have developed a conditional mutant of SARS-CoV-2 with luciferase and fluorescent reporters, which can be safely used under biosafety level 2 conditions. Our single-cycle infectious SARS-CoV-2 virus replicon system can serve as a versatile platform to study SARS-CoV-2 intracellular biology and to perform high-throughput screening of antiviral drugs under BSL2 containment.

Topics & Concepts

BiologyLuciferaseVesicular stomatitis virusRepliconVirologyGreen fluorescent proteinVirusTransfectionPlasmidViral replicationReporter geneViral envelopeVirus-like particleVector (molecular biology)Reverse geneticsCell cultureMicrocarrierInfectious disease (medical specialty)CoronavirusGene expressionViral vectorViral entryPorcine circovirusGlycoproteinMolecular biologyRetrovirusGeneCell biologyMicrobiologySARS-CoV-2 and COVID-19 ResearchSARS-CoV-2 detection and testingInfection Control and Ventilation
Development of a Single-Cycle Infectious SARS-CoV-2 Virus Replicon Particle System for Use in Biosafety Level 2 Laboratories | Litcius