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

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

Molecular virology tools are critical for basic studies of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and for developing new therapeutics. There remains a need for experimental systems that do not rely on viruses capable of spread that could potentially be used in lower containment settings. Here, we develop spike-deleted SARS-CoV-2 self-replicating RNAs using a yeast-based reverse genetics system. These non-infectious self-replicating RNAs, or replicons, can be trans-complemented with viral glycoproteins to generate Replicon Delivery Particles (RDPs) 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 characterizing viral variants.