Live cell assay to quantify the activity of SARS-CoV-2 main protease, Mpro

Applications

• Screening, characterization, and development of protease inhibitors against SARS-CoV-2 and related coronaviruses

Key Benefits & Differentiators

• Robust quantification system: live cell, fluorescence-based gain-of-function assay for inhibitor screening
• High specificity and sensitivity: exquisite specificity and high signal-to-noise ratio

Technology Summary

Despite the ongoing COVID-19 pandemic, there are still no effective drugs to treat disease caused by SARS-CoV-2. SARS-CoV-2 viral proteases (M^pro or PL^pro) and the RNA dependent RNA polymerase (RdRp) are ideal targets for drug discovery. The main protease, M^pro, is required to cleave the viral polyprotein into precise functional units for virus replication and pathogenesis and is therefore an attractive drug target. A wide range of biochemical assays are available for measuring SARS-CoV-2 protease activity, but specific and sensitive cellular assays are less developed. There are currently no sensitive and specific live cell assays available for quantifying SARS-CoV-2 M^pro inhibition, which hinders development of drugs to target this enzyme.

Researchers at the University of Minnesota have developed a quantitative reporter for M^pro function in living cells (Src-Mpro-Tat-eGFP), in which protease inhibition by genetic or chemical methods results in strong eGFP fluorescence (Fig. 1). This robust gain-of-function system readily distinguishes between inhibitor potencies and can be scaled-up to high-throughput platforms for drug development. Essentially, the better the chemical inhibitor/drug, the stronger the fluorescent readout of the assay which expedites any high-throughput readout.

Phase of Development

Desired Partnerships

• License
• Sponsored research
• Co-development

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Researchers

• Reuben S. Harris, PhD Professor, Biochemistry, Molecular Biology, and Biophysics Department