Gain-of-Signal Assays for Probing Inhibition of SARS-CoV-2 M^pro/3CL^pro in Living Cells.

ABSTRACT: The main protease, M^pro, of SARS-CoV-2 is required to cleave the viral polyprotein into precise functional units for virus replication and pathogenesis. Here, we report quantitative reporters for M^pro function in living cells in which protease inhibition by genetic or chemical methods results in robust signal readouts by fluorescence (enhanced green fluorescent protein [eGFP]) or bioluminescence (firefly luciferase). These gain-of-signal systems are scalable to high-throughput platforms for quantitative discrimination between M^pro mutants and/or inhibitor potencies as evidenced by validation of several reported inhibitors. Additional utility is shown by single M^pro amino acid variants and structural information combining to demonstrate that both

SUBMITTER: Moghadasi SA 

PROVIDER: S-EPMC9239272 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature