Gain-of-function assay for SARS-CoV-2 Mpro inhibition in living cells

Seyad Arad Moghadasi, View ORCID ProfileJordan T. Becker, Christopher Belica, Chloe Wick, William L. Brown, View ORCID ProfileReuben S. Harris


The main protease, Mpro, of SARS-CoV-2 is required to cleave the viral polyprotein into precise functional units for virus replication and pathogenesis. Here we demonstrate a quantitative reporter for Mpro function in living cells, in which protease inhibition by genetic or chemical methods results in strong eGFP fluorescence. This robust gain-of-function system readily distinguishes between inhibitor potencies and can be scaled-up to high-throughput platforms for drug testing.