Characterization of a bicistronic knock-in reporter mouse model for investigating the role of CABLES2 in vivo.

Experimental animals 2021 Feb 06

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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 (Cables2Tom) 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 Cables2Tom/Tom revealed interaction of CABLES2 with CDK5. Collectively, our new Cables2 knock-in reporter model will enable the comprehensive analysis of in vivo CABLES2 function.

Citation

Ammar Shaker Hamed Hasan, Tra Thi Huong Dinh, Hoai Thu Le, Saori Mizuno-Iijima, Yoko Daitoku, Miyuki Ishida, Yoko Tanimoto, Kanako Kato, Atsushi Yoshiki, Kazuya Murata, Seiya Mizuno, Fumihiro Sugiyama. Characterization of a bicistronic knock-in reporter mouse model for investigating the role of CABLES2 in vivo. Experimental animals. 2021 Feb 06;70(1):22-30