Generation and Characterization of Ins1-cre-driver C57BL/6N for Exclusive Pancreatic Beta Cell-specific Cre-loxP Recombination

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Author(s)

    • Yoshiki Atsushi
    • Experimental Animal Division, RIKEN BioResource Center, 3-1-1 Koyadai, Tsukuba 305-0074, Japan
    • Takahashi Satoru
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan
    • Sugiyama Fumihiro
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan
    • Yagami Ken-ichi
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan
    • Daitoku Yoko
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan
    • Mizuno Seiya
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan
    • Tanimoto Yoko
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan
    • Mizuno-Iijima Saori
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan
    • Matsuo Miki
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan
    • Kajiwara Noriko
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan
    • Ema Masatsugu
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan|Present address: Research Center for Animal Life Science, Shiga University of Medical Science, Seta, Ootsu, Siga 520-2192, Japan
    • Oishi Hisashi
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan

Abstract

Cre/loxP system-mediated site-specific recombination is utilized to study gene function <i>in vivo</i>. Successful conditional knockout of genes of interest is dependent on the availability of Cre-driver mice. We produced and characterized pancreatic β cell-specific Cre-driver mice for use in diabetes mellitus research. The gene encoding Cre was inserted into the second exon of mouse <i>Ins1</i> in a bacterial artificial chromosome (BAC). Five founder mice were produced by microinjection of linearized <i>BAC Ins1-cre</i>. The transgene was integrated between <i>Mafa</i> and the telomere on chromosome 15 in one of the founders, BAC Ins1-cre25. To investigate Cre-loxP recombination, BAC Ins1-cre25 males were crossed with two different Cre-reporters, R26R and R26GRR females. On gross observation, reporter signal after Cre-loxP recombination was detected exclusively in the adult pancreatic islets in both F<sub>1</sub> mice. Immunohistological analysis indicated that Cre-loxP recombination-mediated reporter signal was colocalized with insulin in pancreatic islet cells of both F<sub>1</sub> mice, but not with glucagon. Moreover, Cre-loxP recombination signal was already observed in the pancreatic islets at E13.5 in both F<sub>1</sub> fetuses. Finally, we investigated ectopic Cre-loxP recombination for <i>Ins1</i>, because the ortholog <i>Ins2</i> is also expressed in the brain, in addition to the pancreas. However, there was no Cre-loxP recombination-mediated reporter signal in the brain of both F<sub>1</sub> mice. Our data suggest that BAC Ins1-cre25 mice are a useful Cre-driver C57BL/6N for pancreatic β cell-specific Cre-loxP recombination, except for crossing with knock-in mice carrying floxed gene on chromosome 15.

Journal

  • Experimental Animals

    Experimental Animals 63(2), 183-191, 2014

    Japanese Association for Laboratory Animal Science

Codes

  • NII Article ID (NAID)
    130003391621
  • NII NACSIS-CAT ID (NCID)
    AA11032321
  • Text Lang
    ENG
  • ISSN
    1341-1357
  • NDL Article ID
    025412043
  • NDL Call No.
    Z54-H752
  • Data Source
    NDL  J-STAGE 
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