We show kinetic dose-response degradation curves for the bromodomain protein, BRD4, endogenously tagged using CRISPR/Cas9 with the peptide HiBiT in HEK293 cells stably expressing the LgBiT fragment. We used the GloMax® Discover microplate luminometer to continuously monitor BRD4 levels over 24 hours after treatment with the BRD4 PROTAC compound MZ1. Rapid degradation of endogenously tagged HiBiT-BRD4 was observed in a dose-dependent fashion with MZ1 PROTAC. At the highest concentration, almost complete degradation of BRD4 was obtained and sustained over the 24-hour period.
Application Note #AN342