| First Author | Murata K | Year | 2021 |
| Journal | J Biochem | Volume | 170 |
| Issue | 4 | Pages | 453-461 |
| PubMed ID | 33982090 | Mgi Jnum | J:386062 |
| Mgi Id | MGI:8345796 | Doi | 10.1093/jb/mvab059 |
| Citation | Murata K, et al. (2021) Efficient induction of proximity-dependent labelling by biotin feeding in BMAL1-BioID knock-in mice. J Biochem 170(4):453-461 |
| abstractText | Proximity-dependent biotin identification (BioID) is a useful method to identify unknown protein-protein interactions. Few reports have described genetically engineered knock-in mouse models for in vivo BioID. Thus, little is known about the proper method for biotin administration and which tissues are applicable. Here, we established a BioID knock-in mouse model of Brain and Muscle ARNT-Like 1 (BMAL1) and the BirA biotin ligase with R118G mutation (BirA*). The BMAL1-BioID mouse model was used to investigate the effect of biotin diet feeding on protein biotinylation in several tissues. The BMAL1-BirA* fusion protein-retained proper intracellular localization of BMAL1 and binding to CLOCK protein in HEK293T cells. A biotin labelling assay in mouse embryonic fibroblasts revealed the protein biotinylation activity of BMAL1-BirA* expressed in knock-in mouse cells depending on biotin supplementation. Lastly, feeding a 0.5% biotin diet for 7 days induced protein biotinylation in the brain, heart, testis and liver of BMAL1-BioID mice without adverse effects on spermatogenesis. In the kidney, the biotin diet increased biotinylated protein levels in BMAL1-BioID and control mice, suggesting the existence of endogenous biotinylation activity. These results provide valuable information to optimize the in vivo BioID procedure. |
