A genetically encoded BRET-based SARS-CoV-2 M^pro protease activity sensor.

ABSTRACT: The main protease, M^pro, is critical for SARS-CoV-2 replication and an appealing target for designing anti-SARS-CoV-2 agents. Therefore, there is a demand for the development of improved sensors to monitor its activity. Here, we report a pair of genetically encoded, bioluminescence resonance energy transfer (BRET)-based sensors for detecting M^pro proteolytic activity in live cells as well as in vitro. The sensors were generated by sandwiching peptides containing the M^pro N-terminal autocleavage sites, either AVLQSGFR (short) or KTSAVLQSGFRKME (long), in between the mNeonGreen and NanoLuc proteins. Co-expression of the sensors with M^pro in live cells resulted in their cleavage while mutation of the critical C145 residue (C145A) in M^pro

SUBMITTER: Geethakumari AM 

PROVIDER: S-EPMC9516532 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature