Science
. 2021 Oct 14;eabj8430.
doi: 10.1126/science.abj8430. Online ahead of print.
Replication and single-cycle delivery of SARS-CoV-2 replicons
Inna Ricardo-Lax # 1 , Joseph M Luna # 1 , Tran Thi Nhu Thao 2 3 4 , Jérémie Le Pen 1 , Yingpu Yu 1 , H-Heinrich Hoffmann 1 , William M Schneider 1 , Brandon S Razooky 1 , Javier Fernandez-Martinez 5 , Fabian Schmidt 6 , Yiska Weisblum 6 , Bettina Salome Trüeb 2 3 , Inês Berenguer Veiga 2 3 , Kimberly Schmied 2 3 , Nadine Ebert 2 3 , Eleftherios Michailidis 1 , Avery Peace 1 , Francisco J Sánchez-Rivera 7 , Scott W Lowe 7 , Michael P Rout 5 , Theodora Hatziioannou 6 , Paul D Bieniasz 6 , John T Poirier 8 , Margaret R MacDonald 1 , Volker Thiel 2 3 , Charles M Rice 1
Affiliations
- PMID: 34648371
- DOI: 10.1126/science.abj8430
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.