Establishment of mouse blastocyst complementation without contribution to germ cells

• Project/Area Number: 15K14374
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Laboratory animal science
• Research Institution: Central Institute for Experimental Animals
• Principal Investigator: Hashimoto Haruo 公益財団法人実験動物中央研究所, 実験動物研究部, 研究員 (30353478)
• Research Collaborator: YUKI YOSHIMURA 公益財団法人実験動物中央研究所, 実験動物研究部, 研究員 ; TOMOO ETOH 公益財団法人実験動物中央研究所, 実験動物研究部, 室長 ; KENJI KAWAI 公益財団法人実験動物中央研究所, 病理解析センター, センター長 ; YUICHIRO HIGUCHI 公益財団法人実験動物中央研究所, 実験動物研究部, 研究員
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 胚盤胞補完法 / 生殖細胞 / PRDM14 / マウス / 不妊 / ES細胞 / キメラマウス / Prdm14

Outline of Final Research Achievements

Our goal is to establish the mouse blastocyst complementation method without contribution to germ cells, and to apply the method to development of humanized pancreatic mice. At first, we established Prdm14 gene deficient ES cells with GFP expression (Prdm14KO-ES cells) using Genome edition technology (CRISPR/Cas9). Then, chimeric mice were generated by Prdm14KO-ES cells and moue blastocysts from IQI/Jic. As the results, the generated chimeric mice were not fertilized. Additionally, Prdm14KO-ES cells did not contribute to germ cells because GFP expression as marker of Prdm14KO-ES cells was not detected in germ cells.
Therefore, these results may solve the problem associated with animal-human chimeric embryo.

Research Products

• [Journal Article] Study on AAV-mediated Gene Therapy for Diabetes in Humanized Liver Mouse to Predict Efficacy in Humans. (2016)
  • Author(s): Hashimoto H, Mizushima T, Ogura T, Kagawa T, Tomiyama K, Takahashi R, Yagoto M, Kawai K, Chijiwa T, Nakamura M, Suemizu H
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 478 Issue: 3 Pages: 1254-60
  • DOI: 10.1016/j.bbrc.2016.08.104
  • Peer Reviewed / Open Access
• [Journal Article] ビニールアイソレータの圧力変化を応用した空気漏洩の簡便な検出方法の確立 (2016)
  • Author(s): 齊藤宗雄, 今井都秦, 橋本晴夫
  • Journal Title: 実験動物技術 Volume: 51 Pages: 41-46
  • Peer Reviewed
• [Journal Article] Expressin of pancreatic and duodenal homeobox1 (PDX1) protein in the interior and exterior regions of the intestine, revealed by development and analysis of Pdx1 knockout mice. (2015)
  • Author(s): Haruo HASHIMOTO, Tsutomu KAMISAKO, Takahiro KAGAWA, Seiki HARAGUCHI, Mika YAGOTO, Ri-ichi TAKAHASHI, Kenji KAWAI, Hiroshi SUEMIZU.
  • Journal Title: Laboratory Animal Research Volume: 31 Issue: 2 Pages: 93-98
  • DOI: 10.5625/lar.2015.31.2.93
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] スンクスの嘔吐を強化する遺伝的要因は雄に存在する可能性. (2015)
  • Author(s): 江袋 進, 橋本晴夫, 冨澤政史, 何 裕遙, 小倉智幸, 樋口裕一郎, 桑畑 玲, 寺田 節, 日置恭司, 高橋利一, 齋藤宗雄
  • Journal Title: 実験動物技術 Volume: 50 Pages: 11-14
  • Peer Reviewed
• [Presentation] マウス雌雄生みわけに関する新技法の開発 (2016)
  • Author(s): 橋本晴夫, 江藤智生, 末水洋志, 伊藤 守
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィッコ横浜
  • Year and Date: 2016-11-30
• [Presentation] C57BL/6JJclを遺伝的背景に持つIRS2欠損マウスの効率的な繁殖方法の検討 (2016)
  • Author(s): 橋本晴夫, 江藤智生, 上迫 努, 山内敏正, 窪田直人, 植木浩二郎, 日置恭司, 齊藤宗雄, 門脇 孝, 伊藤 守
  • Organizer: 第63回日本実験動物学会総会
  • Place of Presentation: ミューザ川崎
  • Year and Date: 2016-05-18
• [Presentation] 2型糖尿病モデルIRS-2ノックアウトマウスにおける遺伝的背景129+Ter/SvおよびC57BL/6JJcl系統の比較 (2015)
  • Author(s): 橋本晴夫、新井敏郎、川井健司、窪田直人、植木浩二郎、日置恭司、斎藤宗雄、大西保行、上山義人、門脇 孝、伊藤 守
  • Organizer: 第 49回日本実験動物技術者協会総会(第34回日本実験動物技術者協会 研究奨励賞)
  • Place of Presentation: 静岡県コンベンションアーツセンター
  • Year and Date: 2015-10-09
  • Invited
• [Presentation] PDX-1ノックアウトマウス開発の試み (2015)
  • Author(s): 橋本晴夫、原口清輝、上迫 努、香川貴洋、位高美香、高橋利一、 川井健司、 末水洋志
  • Organizer: 第62回日本実験動物学会総会
  • Place of Presentation: 京都テルサ
  • Year and Date: 2015-05-28