Induction of high quality induced hepatocytes utilizing the human artificial chromosome

• Project/Area Number: 24790696
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Gastroenterology
• Research Institution: Tokyo Metropolitan Institute of Medical Science
• Principal Investigator: SUZUKI Teruhiko 公益財団法人東京都医学総合研究所, 生体分子先端研究分野, 主席研究員 (70621027)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 染色体 / ヒト人工染色体 / リプログラミング / インテグラーゼ / リコンビナーゼ / 人工染色体 / HAC / 肝細胞 / ダイレクトリプログラミング

Outline of Final Research Achievements

To enable highly efficient loading of multiple gene expression units to the human artificial chromosome (HAC), I developed Simultaneous/sequential Integration of Multivector system (SIM system). This system allows for simultaneous integration of maximum three gene-loading vectors carrying genes of interest by co-transfecting recombinase/integrase expression vectors. This system is useful to construct the HACs for cell reprogramming and monitoring of cell differentiation.

Research Products

• [Journal Article] A novel system for simultaneous or sequential integration of multiple gene-loading vectors into a defined site of a human artificial chromosome. (2014)
  • Author(s): Teruhiko Suzuki, Yasuhiro Kazuki, Mitsuo Oshimura, and Takahiko Hara
  • Journal Title: PLoS One Volume: 9 Issue: 10 Pages: e110404-e110404
  • DOI: 10.1371/journal.pone.0110404
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Development of a new method for the simultaneous integration of multiple vectors into human/mouse artificial chromosome. (2014)
  • Author(s): Teruhiko Suzuki, Manami Kawaguchi, Mitsuo Oshimura, Takahiko Hara
  • Organizer: 第１２回幹細胞シンポジウム
  • Place of Presentation: 九州大学・百年講堂 （福岡）
  • Year and Date: 2014-05-30 – 2014-05-31
• [Presentation] リコンビナーゼ/インテグラーゼを利用してヒト人工染色体に複数ベクターを同時導入する方法の開発 (2013)
  • Author(s): 鈴木輝彦, 押村光雄, 原孝彦
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸
• [Presentation] ダイレクトリプログラミングにより作出したiHepクローンの遺伝子発現解析 (2013)
  • Author(s): 高橋彩実, 鈴木輝彦, 原孝彦
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸
• [Presentation] iPS細胞とヒト/マウス人工染色体を用いた成人発症II型シトルリン血症遺伝子治療モデルの開発 (2012)
  • Author(s): 鈴木輝彦, 宇野愛海, 香月康宏, 黒田英志, 牛飼美晴, 平塚正治, 佐伯武頼, 押村光雄.
  • Organizer: 第11回日本再生医療学会総会
  • Place of Presentation: 横浜
• [Presentation] Towards a non-immunogenic gene delivery vector system using human artificial chromosome for gene- and cell-therapy. (2012)
  • Author(s): Narumi Uno, Teruhiko Suzuki, Kana Ueda, Katsuhiro Uno, Sayaka Fukuhara, Yasuhiro Kazuki, Masaharu Hiratsuka and Mitsuo Oshimura
  • Organizer: ISSCR 10th Annual Meeting.
  • Place of Presentation: 横浜
• [Presentation] Generation of integration-free mouse and human iPS cells using human artificial chromosomes. (2012)
  • Author(s): Ueda, K, Hiratsuka, M, Uno, N, Kurosaki, H, Imaoka, N, Uno, K, Suzuki, T, Kazuki, K, Akakura, Y, Katoh, M, Osaki, M, Kazuki, Y, Oshimura, M.
  • Organizer: ISSCR 10th Annual Meeting.
  • Place of Presentation: 横浜
• [Presentation] 人工染色体を用いたiPS細胞の未分化細胞除去及び腫瘍化抑制システムの開発 (2012)
  • Author(s): 宇野愛海, 宇野勝洋, 上田佳奈, 福原早也佳, 平塚正治, 香月康弘, 鈴木輝彦, 押村光雄
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡
• [Remarks] 公益財団法人東京都医学総合研究所・幹細胞プロジェクト ホームページ
  • URL: http://www.igakuken.or.jp/stem-cell/