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 (October 2021) Replication and single-cycle delivery of SARS-CoV-2 replicons. Science. ISSN 0036-8075
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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. 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.
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