Application Example
Endogenously expressed proteins tagged with HiBiT using CRISPR/CAS9 gene editing can also be used as the BRET energy donor when LgBiT is also expressed in the cell. The HaloTag® fusion can be introduced transiently allowing for interactions kinetics to be assessed under native expression conditions.
Here is an example of an endogenously tagged protein, HiBiT-BRD4, with varying kinetic increases in ubiquitination levels that depend on specific PROTAC treatment.