1. Replication and single-cycle delivery of SARS-CoV-2 replicons.
- Author
-
Ricardo-Lax I, Luna JM, Thao TTN, Le Pen J, Yu Y, Hoffmann HH, Schneider WM, Razooky BS, Fernandez-Martinez J, Schmidt F, Weisblum Y, Trüeb BS, Berenguer Veiga I, Schmied K, Ebert N, Michailidis E, Peace A, Sánchez-Rivera FJ, Lowe SW, Rout MP, Hatziioannou T, Bieniasz PD, Poirier JT, MacDonald MR, Thiel V, and Rice CM
- Subjects
- Animals, Antibodies, Neutralizing immunology, Antibodies, Viral immunology, Antiviral Agents pharmacology, Cell Line, Humans, Interferons pharmacology, Microbial Sensitivity Tests, Mutation, Plasmids, RNA, Viral metabolism, Replicon genetics, Reverse Genetics, SARS-CoV-2 drug effects, SARS-CoV-2 physiology, Saccharomyces cerevisiae genetics, Spike Glycoprotein, Coronavirus genetics, Viral Nonstructural Proteins genetics, Viral Nonstructural Proteins metabolism, Viral Pseudotyping, Virion genetics, Virion physiology, Virus Replication, RNA, Viral genetics, Replicon physiology, SARS-CoV-2 genetics
- 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.
- Published
- 2021
- Full Text
- View/download PDF