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

• Hasegawa Yoshikazu

  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

• 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

Bibliographic Information

Other Title

• Generation of CRISPR/Cas9-mediated bicistronic knock-in ins1-cre driver mice

Abstract

In the present study, we generated novel cre driver mice for gene manipulation in pancreatic β cells. Using the CRISPR/Cas9 system, stop codon sequences of Ins1 were targeted for insertion of cre, including 2A sequences. A founder of C57BL/6J-Ins1^{em1 (cre) Utr} strain was produced from an oocyte injected with pX330 containing the sequences encoding gRNA and Cas9 and a DNA donor plasmid carrying 2A-cre. (R26GRR x C57BL/6J-Ins1^{em1 (cre) Utr}) F₁ 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-Ins1^{em1 (cre) Utr} is useful for studies of glucose metabolism and the strategy of bicistronic cre knock-in using the CRISPR/Cas9 system could be useful for production of cre driver mice.

Journal

• Experimental Animals

  Experimental Animals 65 (3), 319-327, 2016

  Japanese Association for Laboratory Animal Science

Keywords

• bicistronic
• cre-loxP
• CRISPR/Cas9
• pancreatic β cells
• mouse

Details

• CRID: 1390282680022166784
• NII Article ID: 130006882371
• NII Book ID: AA11032321
• DOI: 10.1538/expanim.16-0016
• ISSN: 18817122
• ISSN: 00075124
• ISSN: 13411357
• HANDLE: 2241/00143851
• NDL BIB ID: 027495409
• PubMed: 27053096
• Web Site: https://tsukuba.repo.nii.ac.jp/records/39184
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I027495409
• Web Site: https://www.jstage.jst.go.jp/article/expanim/65/3/65_16-0016/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/2016323287
• Text Lang: en
• Data Source:
  • JaLC
  • IRDB
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed