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. 2021 Jul 21;12(8):1302–1307. doi: 10.1021/acsmedchemlett.1c00285

Figure 5.

Figure 5

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).