Replication and single-cycle delivery of SARS-CoV-2 replicons
Inna Ricardo-Lax, Joseph M. Luna, Tran Thi Nhu Thao, Jérémie Le Pen, Yingpu Yu, Hans-Heinrich Hoffmann, William M. Schneider, Brandon S. Razooky, Javier Fernández-Martı́nez, Fabian Schmidt, Yiska Weisblum, Bettina Salome Trüeb, Inês Berenguer Veiga, Kimberly Schmied, Nadine Ebert, Eleftherios Michailidis, Avery Peace, Francisco J. Sánchez‐Rivera, Scott W. Lowe, Michael P. Rout, Théodora Hatziioannou, Paul D. Bieniasz, John T. Poirier, Margaret R. MacDonald, Volker Thiel, Charles M. Rice
Abstract
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.