Generation of CRISPR/Cas9-mediated bicistronic knock-in <i>ins1-cre</i> driver mice

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著者

    • Hasegawa Yoshikazu
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
    • Yoshiki Atsushi
    • Experimental Animal Division, RIKEN BioResource Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan
    • Yagami Ken-ichi
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
    • Iseki Hiroyoshi
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan|International Institute for Integrative Sleep Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
    • Mizuno Seiya
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
    • Sugiyama Fumihiro
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
    • Hoshino Yoshikazu
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan|Hoshino Laboratory Animals, Inc., 1405 Kouda, Bando, Ibaraki 306-0606, Japan
    • Ibrahim Abdelaziz E.
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan|Department of Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt
    • Kato Kanako
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
    • Daitoku Yoko
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
    • Tanimoto Yoko
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
    • Ikeda Yoshihisa
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan|Charles River Laboratories Japan, Inc., 955 Kamibayashi, Ishioka, Ibaraki 315-0138, Japan
    • Oishi Hisashi
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
    • Takahashi Satoru
    • Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan|International Institute for Integrative Sleep Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan

抄録

In the present study, we generated novel <i>cre</i> driver mice for gene manipulation in pancreatic β cells. Using the CRISPR/Cas9 system, stop codon sequences of <i>Ins1</i> were targeted for insertion of <i>cre</i>, including <i>2A</i> sequences. A founder of C57BL/6J-<i>Ins1<sup>em1 (cre) Utr</sup></i> strain was produced from an oocyte injected with <i>pX330</i> containing the sequences encoding gRNA and Cas9 and a DNA donor plasmid carrying <i>2A-cre</i>. (R26GRR x C57BL/6J-<i>Ins1<sup>em1 (cre) Utr</sup></i>) F<sub>1</sub> mice were histologically characterized for cre-loxP recombination in the embryonic and adult stages; cre-loxP recombination was observed in all pancreatic islets examined in which almost all insulin-positive cells showed tdsRed fluorescence, suggesting β cell-specific recombination. Furthermore, there were no significant differences in results of glucose tolerance test among genotypes (homo/hetero/wild). Taken together, these observations indicated that C57BL/6J-<i>Ins1<sup>em1 (cre) Utr</sup></i> is useful for studies of glucose metabolism and the strategy of bicistronic <i>cre</i> knock-in using the CRISPR/Cas9 system could be useful for production of <i>cre</i> driver mice.

収録刊行物

  • 実験動物彙報

    実験動物彙報 65(3), 319-327, 2016

    公益社団法人 日本実験動物学会

各種コード

  • NII論文ID(NAID)
    130006882371
  • 本文言語コード
    ENG
  • ISSN
    1341-1357
  • データ提供元
    J-STAGE 
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