Naruse Chie, Ishibashi Ojiro, Ohgushi Masatoshi, Imai Hirohiko, Matsuzaki Tomoko, Pan Xuchi, Miyazaki Tatsuhiko, Shidahara Yuka, Shirakawa Yu, Sugiyama Fumihiro, Asano Masahide
Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan.
Laboratory of Developmental Systems, Department of Biosystems Science, Institute for Life and Medical Sciences, Kyoto University, Kyoto 606-8507, Japan.
iScience. 2025 Jun 23;28(7):112992. doi: 10.1016/j.isci.2025.112992. eCollection 2025 Jul 18.
Protein knockdown using a zinc finger degron tag and thalidomide analogs was previously considered ineffective in mouse cells. However, using EGFP as an indicator, we found that a super-degron tag (SD) enables degradation in mouse cells when combined with iberdomide or mezigdomide. While SD-tagged EGFP was degraded in wild-type mouse cells, SD-tagged PD-1 required human-type CEREBLON (CRBN) for degradation. In mice with CRBN, endogenous PD-1 tagged with SD was efficiently degraded in T cells both and . Compared with anti-PD-1 antibody treatment, the degradation of PD-1 led to more rapid activation of CD8 T cells. Moreover, pomalidomide that crosses the brain-blood barrier reduced PD-1 expression in the brain. These results suggest that SD and thalidomide analogs can be used for and protein knockdown in mice, although some conditional settings are required.
此前认为，使用锌指降解标签和沙利度胺类似物进行蛋白质敲低在小鼠细胞中无效。然而，以增强绿色荧光蛋白（EGFP）作为指标，我们发现，一种超级降解标签（SD）与泊马度胺或米度胺联合使用时能够在小鼠细胞中实现降解。虽然带有SD标签的EGFP在野生型小鼠细胞中被降解，但带有SD标签的程序性死亡受体1（PD-1）需要人源型小脑萎缩相关蛋白（CRBN）才能降解。在表达CRBN的小鼠中，带有SD标签的内源性PD-1在T细胞中无论在体内还是体外均被有效降解。与抗PD-1抗体治疗相比，PD-1的降解导致CD8 T细胞更快速地激活。此外，能够穿过血脑屏障的泊马度胺降低了大脑中PD-1的表达。这些结果表明，尽管需要一些条件设置，但SD和沙利度胺类似物可用于小鼠体内和体外的蛋白质敲低。
