Laboratory Animal Resource Center, Trans-Border Medical Research Center, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
Doctor's Program in Biomedical Sciences, Graduate School of Comprehensive Human Science, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
Exp Anim. 2021 Feb 6;70(1):22-30. doi: 10.1538/expanim.20-0063. Epub 2020 Aug 11.
Two members of the CDK5 and ABL enzyme substrate (CABLES) family, CABLES1 and CABLES2, share a highly homologous C-terminus. They interact and associate with cyclin-dependent kinase 3 (CDK3), CDK5, and c-ABL. CABLES1 mediates tumor suppression, regulates cell proliferation, and prevents protein degradation. Although Cables2 is ubiquitously expressed in adult mouse tissues at RNA level, the role of CABLES2 in vivo remains unknown. Here, we generated bicistronic Cables2 knock-in reporter mice that expressed CABLES2 tagged with 3×FLAG and 2A-mediated fluorescent reporter tdTomato. Cables2-3×FLAG-2A-tdTomato (Cables2) mice confirmed the expression of Cables2 in various mouse tissues. Interestingly, high intensity of tdTomato fluorescence was observed in the brain, testis and ovary, especially in the corpus luteum. Furthermore, immunoprecipitation analysis using the brain and testis in Cables2 revealed interaction of CABLES2 with CDK5. Collectively, our new Cables2 knock-in reporter model will enable the comprehensive analysis of in vivo CABLES2 function.
两个细胞周期蛋白依赖性激酶 5 和 ABL 酶底物（CABLES）家族成员，CABLES1 和 CABLES2，具有高度同源的 C 末端。它们与细胞周期蛋白依赖性激酶 3（CDK3）、CDK5 和 c-ABL 相互作用和关联。CABLES1 介导肿瘤抑制，调节细胞增殖，并防止蛋白质降解。尽管 Cables2 在成年小鼠组织中的 RNA 水平上广泛表达，但 Cables2 在体内的作用仍然未知。在这里，我们生成了双顺反子 Cables2 敲入报告小鼠，该小鼠表达了带有 3×FLAG 和 2A 介导的荧光报告 tdTomato 的 CABLES2 标签。Cables2-3×FLAG-2A-tdTomato（Cables2）小鼠证实了 Cables2 在各种小鼠组织中的表达。有趣的是，在大脑、睾丸和卵巢中观察到 tdTomato 荧光强度很高，特别是在黄体中。此外，使用 Cables2 的大脑和睾丸进行免疫沉淀分析表明 CABLES2 与 CDK5 相互作用。总之，我们新的 Cables2 敲入报告模型将能够全面分析体内 CABLES2 功能。
