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Development of protein knockdown method in zygote

Research Project

Project/Area Number 26640063
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Laboratory animal science
Research InstitutionNagahama Institute of Bio-Science and Technology

Principal Investigator

Nakamura Toshinobu  長浜バイオ大学, バイオサイエンス学部, 准教授 (80403202)

Project Period (FY) 2014-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2015: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Keywordsタンパク質ノックダウン / オートファジー / 母性タンパク質 / 受精卵
Outline of Final Research Achievements

In this study, we try to develop a novel method for protein knockdown. We produced chimeric protein consisting of lysosome binding peptide and protein A. When this fusion proteins were microinjected to zygote with EGFP mRNA and anti-EGFP antibody, EGFP proteins were efficiently degraded. Next, we examined whether this method are applicable to endogenous Stella proteins, a maternal proteins localized in nucleus. However, Stella proteins were not degraded after microinjected with a fusion proteins and anti-Stella antibody. These data suggested that this method is applicable to cytoplasmic proteins, but not to nuclear proteins.

Report

(3 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • Research Products

    (45 results)

All 2016 2015 2014 Other

All Journal Article (10 results) (of which Peer Reviewed: 10 results,  Open Access: 9 results,  Acknowledgement Compliant: 7 results) Presentation (26 results) (of which Int'l Joint Research: 3 results,  Invited: 5 results) Book (5 results) Remarks (4 results)

  • [Journal Article] Trichostatin A specifically improves the aberrant expression of transcription factor genes in embryos produced by somatic cell nuclear transfer.2015

    • Author(s)
      Inoue K, Oikawa M, Kamimura S, Ogonuki N, Nakamura T, Nakano T, Abe K, Ogura A.
    • Journal Title

      Sci Rep

      Volume: 5 Issue: 1 Pages: 10127-10127

    • DOI

      10.1038/srep10127

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Human Dynactin-Associated Protein Transforms NIH3T3 Cells to Generate Highly Vascularized Tumors with Weak Cell-Cell Interaction.2015

    • Author(s)
      Kunoh T, Wang W, Kobayashi H, Matsuzaki D, Togo Y, Tokuyama M, Hosoi M, Koseki K, Wada S, Nagai N, Nakamura T, Nomura S, Hasegawa M, Sasaki R, Mizukami T.
    • Journal Title

      PLoS One.

      Volume: 10(8) Issue: 8 Pages: e0135836-e0135836

    • DOI

      10.1371/journal.pone.0135836

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Stella controls chromocenter formation through regulation of Daxx expression in 2-cell embryos.2015

    • Author(s)
      Arakawa, T., Nakatani, T., Oda, M., Kimura, Y., Sekita, Y., Kimura, T., Nakamura, T., and Nakano, T.
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 466 Issue: 1 Pages: 60-60

    • DOI

      10.1016/j.bbrc.2015.08.106

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] De novo DNA methylation through the 5'-segment of the H19 ICR maintains its imprint during early embryogenesis2015

    • Author(s)
      Matsuzaki, H., Okamura, E., Takahashi, T., Ushiki, A., Nakamura, T., Nakano, T., Hata, K., Fukamizu, A., and Tanimoto, K.
    • Journal Title

      Development

      Volume: 142 Pages: 3833-3844

    • DOI

      10.1242/dev.126003

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Global DNA hypomethylation coupled to cellular transformation and metastatic ability.2015

    • Author(s)
      Funaki S, Nakamura T, Nakatani T, Umehara H, Nakashima H, Okumura M, Oboki K, Matsumoto K, Saito H, Nakano T.
    • Journal Title

      FEBS lett.

      Volume: 589 Issue: 24PartB Pages: 4053-60

    • DOI

      10.1016/j.febslet.2015.11.020

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Biotin-deficient diet induces chromosome misalignment and spindle defects in mouse oocytes.2015

    • Author(s)
      Tsuji A, Nakamura T, Shibata K.
    • Journal Title

      Biosci Biotechnol Biochem.

      Volume: 79 Issue: 2 Pages: 292-9

    • DOI

      10.1080/09168451.2014.968090

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Dppa3 expression is critical for generation of fully-reprogrammed iPS cells and maintenance of Dlk1-Dio3 imprinting.2015

    • Author(s)
      Xu X et al.
    • Journal Title

      Nature Communications

      Volume: 6 Issue: 1 Pages: 6008-6008

    • DOI

      10.1038/ncomms7008

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Stella preserves maternal chromosome integrity by inhibiting 5hmC-dependent gammaH2AX accumulation.2015

    • Author(s)
      Nakatani et al.
    • Journal Title

      EMBO Reports

      Volume: 16 Issue: 5 Pages: 582-589

    • DOI

      10.15252/embr.201439427

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Trichostatin A specifically improves the aberrant expression of transcription factor genes in embryos produced by somatic cell nuclear transfer.2015

    • Author(s)
      Inoue K, Oikawa M, Kamimura S, Ogonuki N, Nakamura T, Nakano T, Abe K, Ogura A.
    • Journal Title

      Sci Rep

      Volume: 未定

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Inhibition of maintenance DNA methylation by Stella.2014

    • Author(s)
      Funaki S, Nakamura T, Nakatani T, Umehara H, Nakashima H, Nakano T.
    • Journal Title

      Biochem Biophys Res Commun.

      Volume: 453 Issue: 3 Pages: 455-60

    • DOI

      10.1016/j.bbrc.2014.09.101

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] Critical function of Klf17 for the zygotic genome activation in mice2016

    • Author(s)
      Yui Goto, Asuka Furuta, Kenji Suzuki, Takashi Kohda, Masahito Ikawa, Toshinobu Nakamura
    • Organizer
      International Symposium on “Epigenome dynamics and regulation in germ cells”
    • Place of Presentation
      Kyoto, Japan
    • Year and Date
      2016-02-17
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 全能性細胞で特異的に発現するKlf17の初期発生における必要性2015

    • Author(s)
      後藤 悠比、鈴木 健士、古田 明日香、幸田 尚、伊川 正人、中村 肇伸
    • Organizer
      BMB2015(第38回日本分子生物学会年会・第88回日本生化学会大会 合同大会)
    • Place of Presentation
      神戸
    • Year and Date
      2015-12-01
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] High-affinity, poly-reactive IgA is required for gut homeostatic maintenance to prevent colitis in mice2015

    • Author(s)
      Shinsaku Okai, Fumihito Usui, Makoto Hasegawa, Toshinobu Nakamura, Kazuya Yamamoto, Eri Nishiyama, Hiroshi Mori, Takuji Yamada, Ken Kurokawa, Tamotsu Kato, Eiji Miyauchi, Hiroshi Ohno, Reiko Shinkura
    • Organizer
      BMB2015(第38回日本分子生物学会年会・第88回日本生化学会大会 合同大会)
    • Place of Presentation
      神戸
    • Year and Date
      2015-12-01
    • Related Report
      2015 Annual Research Report
  • [Presentation] High-affinity, poly-reactive IgA is required for gut homeostatic maintenance to prevent colitis in mice2015

    • Author(s)
      Shinsaku Okai, Fumihito Usui, Makoto Hasegawa, Toshinobu Nakamura, Kazuya Yamamoto, Eri Nishiyama, Hiroshi Mori, Takuji Yamada, Ken Kurokawa, Tamotsu Kato, Eiji Miyauchi, Hiroshi Ohno, Reiko Shinkura
    • Organizer
      第44回日本免疫学会学術集会
    • Place of Presentation
      札幌
    • Year and Date
      2015-11-18
    • Related Report
      2015 Annual Research Report
  • [Presentation] 全能性細胞で特異的に発現する遺伝子群の機能解析2015

    • Author(s)
      中村 肇伸
    • Organizer
      神戸大学農学部 第33回インターゲノミクスセミナー
    • Place of Presentation
      神戸
    • Year and Date
      2015-11-13
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] Vitamin B1栄養が卵子の質におよぼす影響2015

    • Author(s)
      辻 愛、中村 肇伸、柴田 克己
    • Organizer
      第54回 日本栄養食糧学会 近畿支部大会
    • Place of Presentation
      神戸
    • Year and Date
      2015-10-10
    • Related Report
      2015 Annual Research Report
  • [Presentation] Critical functions of totipotent-cell specific genes Klf17 and Btg4 in zygotic reprogramming2015

    • Author(s)
      Kenji Suzuki, Yui Goto, Asuka Furuta, Takashi Kohda, Masahito Ikawa, Toshinobu Nakamura
    • Organizer
      The 40th Naito Conference Epigenetics "From Histone Code to Therapeutic Strategy"
    • Place of Presentation
      Sapporo, Japan
    • Year and Date
      2015-09-15
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 全能性細胞で高発現するBtg4の初期発生における必要性2015

    • Author(s)
      鈴木 健士、後藤 悠比、古田 明日香、伊川 正人、中村 肇伸
    • Organizer
      文部科学省科学研究費補助金 新学術領域研究「生殖細胞のエピゲノムダイナミクスとその制御」第3回若手勉強会
    • Place of Presentation
      静岡県
    • Year and Date
      2015-07-22
    • Related Report
      2015 Annual Research Report
  • [Presentation] 初期胚のクロマチン再構成におけるヒストンシャペロンの役割 ~DNA脱メチル化制御因子Stella欠損 胚をモデルとして~2015

    • Author(s)
      荒川 達彦、中谷 庸寿、小田 昌朗、木村 康義、関田 洋一、木村 透、中村 肇 伸、仲野 徹
    • Organizer
      文部科学省科学研究費補助金 新学術領域研究「生殖細胞のエピゲノムダイナミクスとその制御」第3回若手勉強会
    • Place of Presentation
      静岡県
    • Year and Date
      2015-07-22
    • Related Report
      2015 Annual Research Report
  • [Presentation] マウス受精卵におけるStellaの雌性ゲノム保護とその初期胚発生への重要性2015

    • Author(s)
      中谷 庸寿、山縣 一夫、木村 透、小田 昌朗、中島 啓行、堀 真由子、関田 洋 一、荒川 達彦、中村 肇伸、仲野 徹
    • Organizer
      第9回日本エピジェネティクス研究会年会
    • Place of Presentation
      東京
    • Year and Date
      2015-05-25
    • Related Report
      2015 Annual Research Report
  • [Presentation] Biotin-deficient diet induces abnormal oocyte in mouse2015

    • Author(s)
      Ai Tsuji, Toshinobu Nakamura, Katsumi Shibata
    • Organizer
      12th Asian Congress of Nutrition
    • Place of Presentation
      Yokohama
    • Year and Date
      2015-05-14 – 2015-05-18
    • Related Report
      2014 Research-status Report
  • [Presentation] Biotin-deficient diet induces abnormal oocyte in mouse2015

    • Author(s)
      Ai Tsuji, Toshinobu Nakamura, Katsumi Shibata
    • Organizer
      12th Asian Congress of Nutrition
    • Place of Presentation
      Yokohama, Japan
    • Year and Date
      2015-05-14
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 全能性細胞で高発現する遺伝子群の機能解析2015

    • Author(s)
      中村 肇伸、仲野 徹
    • Organizer
      CREST「エピゲノム研究に基づく診断・治療へ向けた新技術の創出」研究領域 第4回領域会議
    • Place of Presentation
      大阪
    • Year and Date
      2015-02-21 – 2015-02-22
    • Related Report
      2014 Research-status Report
  • [Presentation] エピゲノム成立の分子メカニズムの解明と制御2015

    • Author(s)
      仲野 徹、中村 肇伸
    • Organizer
      CREST「エピゲノム研究に基づく診断・治療へ向けた新技術の創出」研究領域 第4回領域会議
    • Place of Presentation
      大阪
    • Year and Date
      2015-02-21 – 2015-02-22
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] マウスにおけるビオチン欠乏は卵子の質を劣化させる2014

    • Author(s)
      辻 愛、中村 肇伸、柴田 克己
    • Organizer
      第13回日本栄養改善学会近畿支部学術総会
    • Place of Presentation
      京都
    • Year and Date
      2014-12-07
    • Related Report
      2014 Research-status Report
  • [Presentation] Asymmetry in histone H2B O-GlcNAcylation between paternal and maternal chromatin of mouse zygotes2014

    • Author(s)
      中村 肇伸、鈴木 健士、後藤 悠比、中谷 庸寿、関田 洋一、仲野 徹
    • Organizer
      第37回日本分子生物学会年会
    • Place of Presentation
      横浜
    • Year and Date
      2014-11-25 – 2014-11-27
    • Related Report
      2014 Research-status Report
  • [Presentation] 腸炎モデルマウスに対する腸管IgA抗体の作用機序の解明2014

    • Author(s)
      岡井 晋作、臼井 文人、野村 慎太郎、中村 肇伸、山本 和也、西山 依里、森 宙史、山田 拓司、黒川 顕、加藤 保、大野 博司、新蔵 礼子
    • Organizer
      第37回日本分子生物学会年会
    • Place of Presentation
      横浜
    • Year and Date
      2014-11-25 – 2014-11-27
    • Related Report
      2014 Research-status Report
  • [Presentation] 初期胚のクロマチン再構成におけるヒストンシャペロンの役割 ~DNA脱メチル化制御因子Stellaの欠損胚をモデルとして~2014

    • Author(s)
      荒川 達彦、中谷 庸寿、関田 洋一、木村 透、中村 肇伸、小田 昌朗、仲野 徹
    • Organizer
      第37回日本分子生物学会年会
    • Place of Presentation
      横浜
    • Year and Date
      2014-11-25 – 2014-11-27
    • Related Report
      2014 Research-status Report
  • [Presentation] 受精卵における胚性遺伝子の活性化機構の解明2014

    • Author(s)
      中村 肇伸
    • Organizer
      文部科学省科学研究費補助金 新学術領域研究「生殖細胞のエピゲノムダイナミクスとその制御」第2回公開シンポジウム
    • Place of Presentation
      福岡
    • Year and Date
      2014-10-31 – 2014-11-01
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] Asymmetry in histone H2B O-GlcNAcylation between paternal and maternal chromatin of mouse zygotes2014

    • Author(s)
      Toshinobu Nakamura, Kenji Suzuki, Yui gotou, Tsunetoshi Nakatani, Yoichi Sekita, Toru Nakano
    • Organizer
      Cold Spring Harbor Laboratory meeting EPIGENETICS & CHROMATIN
    • Place of Presentation
      New York, USA
    • Year and Date
      2014-09-09 – 2014-09-13
    • Related Report
      2014 Research-status Report
  • [Presentation] Stella preserves maternal chromosome integrity by inhibiting 5hmC-dependent γH2AX accumulation2014

    • Author(s)
      Tsunetoshi Nakatani, Kazuo Yamagata, Tohru Kimura, Masaaki Oda, Hiroyuki Nakashima, Mayuko Hori, Yoichi Sekita, Tatsuhiko Arakawa, Toshinobu Nakamura, Toru Nakano
    • Organizer
      Cold Spring Harbor Laboratory meeting EPIGENETICS & CHROMATIN
    • Place of Presentation
      New York, USA
    • Year and Date
      2014-09-09 – 2014-09-13
    • Related Report
      2014 Research-status Report
  • [Presentation] トリコスタチンA処理による体細胞核移植クローン胚の転写因子関連遺伝子発現の改善について2014

    • Author(s)
      井上 貴美子、及川 真実、上村 悟氏、越後貫 成美、中村 肇伸、仲野 徹、小倉 淳郎
    • Organizer
      第107回日本繁殖生物学会大会
    • Place of Presentation
      帯広
    • Year and Date
      2014-08-21 – 2014-08-24
    • Related Report
      2014 Research-status Report
  • [Presentation] 初期胚のクロマチン再構成におけるヒストンシャペロンの役割 ~DNA脱メチル化制御因子Stellaの欠損胚をモデルとして~2014

    • Author(s)
      荒川 達彦、中谷 庸寿、関田 洋一、木村 透、中村 肇伸、小田 昌朗、仲野 徹
    • Organizer
      第3回大阪大学医学系研究科若手研究フォーラム
    • Place of Presentation
      大阪
    • Year and Date
      2014-07-30
    • Related Report
      2014 Research-status Report
  • [Presentation] 全能性細胞で高発現するBtg4の機能解析2014

    • Author(s)
      鈴木 健士、後藤 悠比、中村 肇伸
    • Organizer
      文部科学省科学研究費補助金 新学術領域研究「生殖細胞のエピゲノムダイナミクスとその制御」第2回若手勉強会
    • Place of Presentation
      つくば
    • Year and Date
      2014-07-16 – 2014-07-18
    • Related Report
      2014 Research-status Report
  • [Presentation] 着床前胚におけるKlf17の重要な役割2014

    • Author(s)
      後藤 悠比、鈴木 健士、中村 肇伸
    • Organizer
      文部科学省科学研究費補助金 新学術領域研究「生殖細胞のエピゲノムダイナミクスとその制御」第2回若手勉強会
    • Place of Presentation
      つくば
    • Year and Date
      2014-07-16 – 2014-07-18
    • Related Report
      2014 Research-status Report
  • [Presentation] 受精卵におけるヒストンH2BのN-アセチルグルコサミン修飾2014

    • Author(s)
      中村 肇伸、鈴木 健士、後藤 悠比、中谷 庸寿、関田 洋一、仲野 徹
    • Organizer
      第8回日本エピジェネティクス研究会年会
    • Place of Presentation
      東京
    • Year and Date
      2014-05-25 – 2014-05-27
    • Related Report
      2014 Research-status Report
  • [Book] 生体の科学「着床前胚と始原生殖細胞におけるDNAメチル化リプログラミング」2015

    • Author(s)
      中村 肇伸
    • Total Pages
      7
    • Publisher
      医学書院
    • Related Report
      2015 Annual Research Report
  • [Book] 細胞工学「母性因子を用いたiPS細胞の樹立効率と質の改善」2015

    • Author(s)
      中村 肇伸
    • Total Pages
      8
    • Publisher
      学研メディカル秀潤社
    • Related Report
      2015 Annual Research Report
  • [Book] 細胞工学; 母性因子を用いたiPS細胞の樹立効率と質の改善2015

    • Author(s)
      中村 肇伸
    • Publisher
      学研メディカル秀潤社
    • Related Report
      2014 Research-status Report
  • [Book] 生体の科学; 着床前胚と始原生殖細胞におけるDNAメチル化リプログラミング2015

    • Author(s)
      中村 肇伸
    • Publisher
      医学書院
    • Related Report
      2014 Research-status Report
  • [Book] Stella and zygotic reprogramming; In “Epigenetic Mechanisms in Cellular Reprogramming”, Epigenetics and Human Health series2015

    • Author(s)
      Nakamura T, Nakano T.
    • Total Pages
      12
    • Publisher
      Springer
    • Related Report
      2014 Research-status Report
  • [Remarks] 中村肇伸准教授らの共同研究が『Nature communications』に掲載されました

    • URL

      http://www.nagahama-i-bio.ac.jp/research/

    • Related Report
      2015 Annual Research Report
  • [Remarks] [プレスリリース]中村肇伸准教授らの共同研究について

    • URL

      http://www.nagahama-i-bio.ac.jp/research/

    • Related Report
      2014 Research-status Report
  • [Remarks] 中村肇伸准教授らの共同研究が『Nature communications』に掲載されました

    • URL

      http://www.nagahama-i-bio.ac.jp/others/

    • Related Report
      2014 Research-status Report
  • [Remarks] Nature communicationsに掲載された論文が新聞で報道されました

    • URL

      http://b-lab.nagahama-i-bio.ac.jp/?page_id=203

    • Related Report
      2014 Research-status Report

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Published: 2014-04-04   Modified: 2017-05-10  

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