HiBiT (top)
and NanoBRET (bottom) target engagement with degraders.
(A) HiBiT, which complements with LgBiT to form the luminescence,
was inserted via CRISPR/Cas9 to the C-terminus of CDK2 in HEK293T
cells. Following treatment with degraders, protein degradation was
monitored by the loss of luminescence. (B) In HEK293T cells, NanoLuc–CDK2
was used as the energy donor and HaloTag–CRBN was used as the
energy acceptor. Following treatment with degraders, the CDK2:CRBN
complex formation was monitored by the NanoBRET ratio (acceptor emission
at 618 nm/donor emission at 460 nm).