Replication and single-cycle delivery of SARS-CoV-2 replicons

• Published in: Science (American Association for the Advancement of Science) Vol. 374; no. 6571; pp. 1099 - 1106
• Main Authors: Ricardo-Lax, Inna, Luna, Joseph M, Thao, Tran Thi Nhu, Le Pen, Jérémie, Yu, Yingpu, Hoffmann, H-Heinrich, Schneider, William M, Razooky, Brandon S, Fernandez-Martinez, Javier, Schmidt, Fabian, Weisblum, Yiska, Trüeb, Bettina Salome, Berenguer Veiga, Inês, Schmied, Kimberly, Ebert, Nadine, Michailidis, Eleftherios, Peace, Avery, Sánchez-Rivera, Francisco J, Lowe, Scott W, Rout, Michael P, Hatziioannou, Theodora, Bieniasz, Paul D, Poirier, John T, MacDonald, Margaret R, Thiel, Volker, Rice, Charles M
• Format: Journal Article
• Language: English
• Published: United States The American Association for the Advancement of Science 26.11.2021
• Subjects: Animals; Antibodies, Neutralizing - immunology; Antibodies, Viral - immunology; Antiviral Agents - pharmacology; Cell Line; Coronaviruses; COVID-19; Drug screening; Genetics; Genomes; Glycoproteins; Humans; Interferons - pharmacology; Microbial Sensitivity Tests; Mutation; Neutralization; Plasmids; Replication; Replicon - genetics; Replicon - physiology; Respiratory diseases; Reverse Genetics; RNA viruses; RNA, Viral - genetics; RNA, Viral - metabolism; Saccharomyces cerevisiae - genetics; SARS-CoV-2 - drug effects; SARS-CoV-2 - genetics; SARS-CoV-2 - physiology; Screening; Severe acute respiratory syndrome; Severe acute respiratory syndrome coronavirus 2; Spike Glycoprotein, Coronavirus - genetics; Spike protein; Viral diseases; Viral Nonstructural Proteins - genetics; Viral Nonstructural Proteins - metabolism; Viral Pseudotyping; Virion - genetics; Virion - physiology; Virus Replication; Viruses; Yeast; Yeasts
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.abj8430

Molecular virology tools are critical for basic studies of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and for developing new therapeutics. Experimental systems that do not rely on viruses capable of spread are needed for potential use in lower-containment settings. In this work, we use a yeast-based reverse genetics system to develop spike-deleted SARS-CoV-2 self-replicating RNAs. These noninfectious self-replicating RNAs, or replicons, can be trans-complemented with viral glycoproteins to generate replicon delivery particles for single-cycle delivery into a range of cell types. This SARS-CoV-2 replicon system represents a convenient and versatile platform for antiviral drug screening, neutralization assays, host factor validation, and viral variant characterization.