核移植クローン技術を用いたゲノム再プログラム化機構の解析

• Project/Area Number: 15080210
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: 小倉 淳郎 The Institute of Physical and Chemical Research, 遺伝工学基盤技術室, 室長 (20194524)
• Project Period (FY): 2003 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥123,200,000 (Direct Cost: ¥123,200,000); Fiscal Year 2007: ¥23,200,000 (Direct Cost: ¥23,200,000); Fiscal Year 2006: ¥23,500,000 (Direct Cost: ¥23,500,000); Fiscal Year 2005: ¥25,500,000 (Direct Cost: ¥25,500,000); Fiscal Year 2004: ¥25,500,000 (Direct Cost: ¥25,500,000); Fiscal Year 2003: ¥25,500,000 (Direct Cost: ¥25,500,000)
• Keywords: 核移植クローン / 生殖細胞 / 胚盤胞 / マウス / X染色体 / 始原生殖細胞 / 体細胞 / 顕微授精 / 胚性遺伝子 / eIF-1A / 幹細胞 / Hdacl / 造血幹細胞 / 胎盤 / NKT細胞 / TS細胞 / 核移植 / 体細胞クローン

Research Abstract

本研究は、胚および産子に異常を生じる体細胞核移植クローン技術を利用することにより体細胞ゲノムと生殖細胞ゲノムの相違が明らかにすることを目的とする。この問題点の理解への近道はおそらく、geneticな要因をできる限り排除してepigeneticな要因に限定でき、ランダムでなく定型的な異常であり、そして発生のできるだけ初期に現れる現象の解析にある。これらの条件を満たす解析対象の一つがマウスの初期胚である。昨年の2-cellに始まるmajor zygotic gene activation(ZGA)の遺伝子発現パターン解析に加えて、今年度は、胚盤胞の遺伝子発現パターンをマイクロアレイ法を用いて解析した。これは、胎盤異常を生じさせる遺伝子を同定する目的である。卵丘細胞クローン、セルトリ細胞クローン、TSA処理卵丘細胞クローン(発生は改善するが、胎盤は異常)、始原生殖細胞クローン、細胞質注入顕微授精(クローンではないが、胎盤は異常)、IVF胚(コントロール)の6種類を比較した。その結果、クローンはX染色体上の遺伝子が特異的に低下することが明らかになり、その中に胎盤発生に重要な遺伝子が含まれていた。興味深いことに、これらの遺伝子のいつくかは細胞質注入顕微授精でも低下しており、胎盤異常との密接な関係が示唆された。逆に、始原生殖細胞クローンは、改善が見られた。解析対象には、胎盤が異常を示さない129系統マウスのクローン、およびES細胞クローンも追加する予定である。今後はその上流にある共通の現象を突き止め、胎盤異常が生じる機構を明らかにしたい。

Research Products

• [Journal Article] Role of retrotransposon-derived imprinted gene, Rtll, in the feto-maternal interface of mouse placenta. (2008)
  • Author(s): Sekita, et. al.
  • Journal Title: Nat. Genet. 40 Pages: 243-248
  • Peer Reviewed
• [Journal Article] The Developmental Ability of Vitrified Oocytes from Different Mouse Strains Assessed by Parthenogenetic Activation and Intracytoplasmic Sperm Injection. (2007)
  • Author(s): Endoh, et. al.
  • Journal Title: J. Reprod. Dev 53 Pages: 1199-1206
  • Peer Reviewed
• [Journal Article] Isolation, characterization, and in vitro and in vivo differentiation of putative thecal stem cells. (2007)
  • Author(s): Honda, et. al.
  • Journal Title: Proc. Natl. Acad. Sci. USA 104 Pages: 12389-12394
  • Peer Reviewed
• [Journal Article] Efficient production of intersubspecific hybrid mice and embryonic stem cells by intracytoplasmic sperm injection. (2007)
  • Author(s): Shinmen A., et al.
  • Journal Title: Molecular Reproduction and Development (in press)
• [Journal Article] Differential developmental ability of embryos cloned from tissue-specific stem cells. (2007)
  • Author(s): Inoue K., et al.
  • Journal Title: Stem Cells (in press)
• [Journal Article] Inefficient reprogramming of the hematopoietic stem cell genome following nuclear transfer. (2006)
  • Author(s): Inoue K., et al.
  • Journal Title: Journal of Cell Science 119 Pages: 1985-1991
• [Journal Article] Spermatozoa and spermatids retrieved from frozen reproductive organs or frozen whole bodies of male mice can produce normal offspring. (2006)
  • Author(s): Ogonuki N., et al.
  • Journal Title: Proceedings of National Academy of Sciences, USA 103 Pages: 13098-13103
• [Journal Article] Chorioallantoic placenta defects in cloned mice. (2006)
  • Author(s): Wakisaka N., et al.
  • Journal Title: Biochem Biophys Res Commun 349 Pages: 106-114
• [Journal Article] Intra-strain polymorphisms are detected but no genomic alteration is found in cloned mice. (2006)
  • Author(s): Gotoh S., et al.
  • Journal Title: Biochem Biophys Res Commun 348 Pages: 166-169
• [Journal Article] Improvement of cumulus-free oocyte maturation in vitro and its application to microinsemination with primary spernmatocytes in mice. (2006)
  • Author(s): Miki H., et al.
  • Journal Title: Journal of Reproduction and Development (印刷中)
• [Journal Article] Microinsemination and nuclear transfer using male germ cells. (2005)
  • Author(s): Ogura A., et al.
  • Journal Title: International Reviews of Cytology 246 Pages: 189-229
• [Journal Article] Generation of cloned mice by direct nuclear transfer from natural killer T Cells. (2005)
  • Author(s): Inoue K., et al.
  • Journal Title: Current Biology 15 Pages: 2005-2005
• [Journal Article] Differential development of rabbit embryos following microinsemination with sperm and spermatids. (2005)
  • Author(s): Ogonuki N., et al.
  • Journal Title: Molecular Reproduction and Development 72 Pages: 411-417
• [Journal Article] Variation in gene expression and aberrantly regulated chromosome regions in cloned mice. (2005)
  • Author(s): Kohda T., et al.
  • Journal Title: Biology of Reproduction 73 Pages: 1302-1311
• [Journal Article] Noninvasive visualization of molecular events in the mammalian zygote. (2005)
  • Author(s): Yamagata K., et al.
  • Journal Title: Genesis 43 Pages: 71-79
• [Journal Article] Birth of offspring after transfer of Mongolian gerbil (Meriones unguiculatus) embryos cryopreserved by vitrification (2005)
  • Author(s): Mochida, K. et al.
  • Journal Title: Molecular Reproduction and Development 70 Pages: 464-470
• [Journal Article] Long-term culture of mouse male germline stem cells under serum- or feeder-free conditions. (2005)
  • Author(s): Kanatsu-Shinohara, M. et al.
  • Journal Title: Biology of Reproduction (in press)
• [Journal Article] Birth of Mice Produced by Gem Cell Nuclear Transfer. (2005)
  • Author(s): Miki, H. et al.
  • Journal Title: Genesis 41 Pages: 81-86
• [Journal Article] Cytoplasmic Asters are Required for Progression past the First Cell Cycle in Cloned Mouse Embryos (2004)
  • Author(s): Miki, H. et al.
  • Journal Title: Biology of Reproduction 71 Pages: 2022-2028
• [Journal Article] Spermatogenesis from epiblast and primordial germ cells following transplantation into postnatal mouse testis (2004)
  • Author(s): Chuma, S. et al.
  • Journal Title: Development 132 Pages: 117-122
• [Journal Article] Generation of pluripotent stem cells from neonatal mouse testis. (2004)
  • Author(s): Kanatsu-Shinohara, M. et al.
  • Journal Title: Cell 119 Pages: 1001-1012
• [Presentation] Characterization of thecal stem cells and primordial oocytes concominantly isolated from mouse neonatal ovaries (2007)
  • Author(s): Ogura, A.
  • Organizer: 4th Annual Conference of Asian Reproductive Biotechnology Society, 2007
  • Place of Presentation: シンガポール
  • Year and Date: 2007-11-24
• [Book] 生殖細胞の発生・エピジェネティクス・再プログラム化(分担執筆) (2007)
  • Author(s): 井上貴美子、小倉淳郎
  • Total Pages: 2224
  • Publisher: 共立出版
• [Book] クロマチン・染色体実験プロトコール(押村光雄, 平岡泰(編)) (2004)
  • Author(s): 井上貴美子, 小倉厚淳郎
  • Total Pages: 231
  • Publisher: 羊土社
• [Remarks]
  • URL: http://www.brc.riken.go.jp/lab/kougaku/
• [Remarks]
  • URL: http://www.riken.jp/r-world/research/lab/brc/engineering/index.html
• [Publications] Fulka-Jr, J., Miyashita, N., Nagai, T., Ogura, A.: "Do cloned mammals skip a reprogramming step?"Nature Biotechnology. 22. 25-26 (2004)
• [Publications] Inoue, K., Ogonuki, N., Mochida, K., Yamamoto, Y., Takano, K., Kohda, T., Ishino, F., Ogura, A.: "Effects of Donor Cell Type and Genotype on the Efficiency of Mouse Somatic Cell Cloning."Biology of Reproduction. 69. 1394-1400 (2003)
• [Publications] Inoue, K., Ogonuki, N., Yamamoto, Y., Takano, K., Miki, H., Mochida, K., Ogura, A.: "Tissue-specific distribution of donor mitochondrial DNA in cloned mice produced by somatic cell nuclear transfer."Genesis. in press. (2004)
• [Publications] Kanatsu-Shinohara, M., Ogonuki, N., Inoue, K., Miki, H., Ogura, A., Toyokuni, S., Shinohara, T.: "Long-Term Proliferation in Culture and Germline Transmission of Mouse Male Germline Stem Cells."Biology of Reproduction. 69. 612-616 (2003)
• [Publications] Kanatsu-Shinohara, M., Ogonuki, N., Inoue, K., Ogura, A., Toyokuni, S., Shinohara, T.: "Restoration of fertility in infertile mice by transplantation of cryopreserved male germline stem cells."Human Reproduction. 18. 2660-2667 (2003)
• [Publications] Miki, H., et al.: "Microinsemination with first-wave round spermatids from immature male mice."Journal of Reproduction and Development. 50. 131-137 (2004)
• [Publications] 小倉淳郎: "これからの大学等研究施設「生命科学編」"文教施設協会. 150 (2003)