A PCR amplicon-based SARS-CoV-2 replicon for antiviral evaluation

Abstract The development of specific antiviral compounds to SARS-CoV-2 is an urgent task. One the obstacles for requirement biocontainment because infectious must be handled in a biosafety level-3 laboratory. Replicon, non-infectious self-replicative viral RNA, could safe and effective tool evaluation. Herein, we generated PCR-based replicon. Eight fragments covering entire genome except S, E, M genes were amplified with HiBiT-tag sequence by PCR. amplicons ligated vitro transcribed RNA. cells electroporated replicon RNA showed more than 3000 times higher luminescence MOCK control at 24 h post-electroporation, indicating robust translation replication was drastically inhibited remdesivir, polymerase inhibitor SARS-CoV-2. IC 50 remdesivir this study 0.29 μM, generally consistent obtained using previous (0.77 μM). Taken together, system applied evaluation without Because transient system, further improvement including establishment stable cell line achieved.