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

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

doi:10.1538/expanim.20-0063

Original

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

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

Author information

Ammar Shaker Hamed Hasan
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
Ministry of Works, Municipalities Affairs and Urban Planning, Building 86, Block 318, Sheikh Hamad Street 1802, Manama Diplomatic Area, Manama, Bahrain
Tra Thi Huong Dinh
Laboratory Animal Resource Center, Trans-Border Medical Research Center, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
Hoai Thu Le
Laboratory Animal Resource Center, Trans-Border Medical Research Center, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
Ph.D. Program in Human Biology, School of Integrative and Global Majors (SIGMA), University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
Saori Mizuno-Iijima
Experimental Animal Division, RIKEN BioResource Research Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan
Yoko Daitoku
Laboratory Animal Resource Center, Trans-Border Medical Research Center, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
Miyuki Ishida
Laboratory Animal Resource Center, Trans-Border Medical Research Center, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
Yoko Tanimoto
Laboratory Animal Resource Center, Trans-Border Medical Research Center, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
Kanako Kato
Laboratory Animal Resource Center, Trans-Border Medical Research Center, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
Atsushi Yoshiki
Experimental Animal Division, RIKEN BioResource Research Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan
Kazuya Murata
Laboratory Animal Resource Center, Trans-Border Medical Research Center, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
Seiya Mizuno
Laboratory Animal Resource Center, Trans-Border Medical Research Center, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
Fumihiro Sugiyama
Laboratory Animal Resource Center, Trans-Border Medical Research Center, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan

Keywords: bicistronic expression system, Cables2, FLAG-tag, knock-in reporter mouse, tdTomato

JOURNAL OPEN ACCESS
Supplementary material

2021 Volume 70 Issue 1 Pages 22-30

DOI https://doi.org/10.1538/expanim.20-0063

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Abstract

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^Tom) 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^Tom/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.

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