Generation of alpha1,3-galactosyltransferase and cytidine monophospho-N-acetylneuraminic acid hydroxylase gene double-knockout pigs

• Project/Area Number: 26870630
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: General surgery; Integrative animal science
• Research Institution: Meiji University
• Principal Investigator: Matsunari Hitomi 明治大学, 研究・知財戦略機構, 特任講師 (70639517)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2015: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 異種移植 / ブタ / ゲノム編集 / 体細胞核移植 / 臓器移植 / 遺伝子ノックアウト / 移植・再生医療 / トランスレーショナルリサーチ

Outline of Final Research Achievements

Pig to human xenotransplantation is one of the current strategies for overcoming the worldwide shortage of organ transplants. However, discordant xenografts involve severe immune rejections strongly related to species-specific glycoantigens. The major xenoantigen is the α-Gal epitope. Another carbohydrate xenoantigen, N-glycolylneuraminic acid (Neu5Gc), which is a component of the H-D antigen, has also been identified. Neu5Gc is synthesized by cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH). The aim of this study was to produce α1,3-galactosyltransferase (GalT)/CMAH-double KO pigs using genome editing and somatic cell nuclear transfer (SCNT) technology. The homozygous CMAH KO cells were established using ZFNs or TALENs. These cells were used for SCNT and produced GalT/CMAH-double KO cloned pigs. The absence of α-Gal and H-D antigens was confirmed in both the tissue and granulocyte samples of the cloned piglets.

Research Products

• [Journal Article] Generation of α1,3-galactosyltransferase and cytidine monophospho-<i>N</i>-acetylneuraminic acid hydroxylase gene double-knockout pigs (2015)
  • Author(s): Miyagawa S, Matsunari H, Watanabe M, Nakano K, Umeyama K, Sakai R, Takayanagi S, Takeishi T, Fukuda T, Yashima S, Maeda A, Eguchi H, Okuyama H, Nagaya M, Nagashima H.
  • Journal Title: Journal of Reproduction and Development Volume: 61 Issue: 5 Pages: 449-457
  • DOI: 10.1262/jrd.2015-058
  • NAID: 130005104116
  • ISSN: 0916-8818, 1348-4400
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Production of alpha 1,3-galactosyltransferase and cytidine monophosphate-N-acetylneuraminic acid hydroxylase gene double-knockout pigs by genome editing and somatic cell cloning (2015)
  • Author(s): Matsunari H, Watanabe M, Nakano K, Uchikura A, Asano Y, Hatae S, Takeishi T, Yashima S, Fukuda T, Sakai S, Umeyama K, Nagaya M, Maeda A, Eguchi H, Okuyama H, Miyagawa S, Nagashima H.
  • Organizer: 2015 IPITA・IXA・CTS JOINT CONGRESS
  • Place of Presentation: Melbourne, Australia
  • Year and Date: 2015-11-15
  • Int'l Joint Research
• [Presentation] α-ガラクトシル抗原およびH-D抗原を発現しない遺伝子ノックアウトブタの作出 (2015)
  • Author(s): 松成ひとみ，渡邊將人，中野和明，内倉鮎子，浅野吉則，武石透輝，福田暢，八島紗耶香，梅山一大，高柳就子，坂井理恵子，前田晃，江口寛，奥山宏臣，宮川周士，長嶋比呂志
  • Organizer: 第17回日本異種移植研究会
  • Place of Presentation: 自治医科大学(栃木県・下野市)
  • Year and Date: 2015-03-14
• [Presentation] Production efficiency of gene knockout pigs using genome editing and somatic cell cloning (2015)
  • Author(s): Hitomi Matsunari, Masahito Watanabe, Kazuaki Nakano, Ayuko Uchikura, Yoshinori Asano, Shota Hatae, Toki Takeishi, Kazuhiro Umeyama, Masaki Nagaya, Shuji Miyagawa, Yutaka Hanazono, Hiromitsu Nakauchi, Hiroshi Nagashima
  • Organizer: 41st Annual Conference of the International Embryo Transfer Society
  • Place of Presentation: Versailles (France)
  • Year and Date: 2015-01-12
• [Remarks] 明治大学バイオリソース研究国際インスティテュート
  • URL: http://muiibr.com
• [Remarks] 明治大学バイオリソース研究国債インスティテュート(MUIIBR)
  • URL: http://muiibr.com