Regulation of intranuclear arrangement of genes and construction of cells which highly express recombinant proteins

• Project/Area Number: 23658091
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied biochemistry
• Research Institution: Mie University
• Principal Investigator: OKUMURA Katsuzumi 三重大学, 生物資源学研究科, 教授 (30177183)
• Project Period (FY): 2011-04-28 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2011: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 動物細胞 / 遺伝子発現 / DNAメチル化 / ヘテロクロマチン / DNA複製 / ゼブラフィッシュ / 核内配置 / エピジェネティクス / DNA損傷 / 核マトリックス

Outline of Final Research Achievements

For the purpose of establishment of cells with efficient expression of recombinant proteins, intranuclear genome dynamics and the genome editing method were investigated. The following results were obtained: The intranuclear arrangement of genes during adipocyte differentiation was not changed even if they expressed. Overexpression of hemimethylated DNA binding protein UHRF1 induced the abnormal shape of the nucleus. Folate deficiency causes the DNA hypomethylation and induces the DNA damage. The specific large genomic region can be deleted by a CRISPR/Cas9 system. Various basic data on intranuclear DNA replication dynamics of Zebrafish cultured cells have been collected.

Research Products

• [Journal Article] 核マトリックスとゲノムダイナミクス (2011)
  • Author(s): 奥村克純
  • Journal Title: 生体の科学 Volume: 62 Pages: 400-401
• [Journal Article] 核内ゲノムダイナミクスの可視化 (2011)
  • Author(s): 奥村克純
  • Journal Title: 生体の科学 Volume: 62 Pages: 500-501
• [Presentation] Molecular and Cellular Analysis Using Zebrafish Derived Cells (2014)
  • Author(s): Kenji Kuriya, Eriko Higashiyama, Eriko Avsal, Yutaka Tamaru, Shin Ogata, and Katsuzumi Okumura
  • Organizer: The Sixth International Workshop on Regional Innovation Studies
  • Place of Presentation: Regional Innovation Hall, Mie University, Tsu, Japan
  • Year and Date: 2014-10-16 – 2014-10-17
• [Presentation] The Lack of the Nutrition Ingredient Induces the DNA Damage (2014)
  • Author(s): Yuta Minamigawa and Katsuzumi Okumura
  • Organizer: The Sixth International Workshop on Regional Innovation Studies
  • Place of Presentation: Regional Innovation Hall, Mie University, Tsu, Japan
  • Year and Date: 2014-10-16 – 2014-10-17
• [Presentation] ゼブラフィッシュ由来細胞を用いたDNA複製の解析 (2014)
  • Author(s): 東山恵理子，栗谷健志，アヴシャル恵理子，田丸浩，緒方進，奥村克純
  • Organizer: 日本農芸化学会中部支部第171回例会
  • Place of Presentation: 名古屋大学シンポジオン(名古屋市千種区)
  • Year and Date: 2014-10-11
• [Presentation] Epigenetic abnormalities: DNA damage induction by DNA hypomethylation and the nutritional connection (2014)
  • Author(s): Katsuzumi Okumura
  • Organizer: The 1st International Symposium for Bioengineering Research and Education
  • Place of Presentation: UCSD, Building 1, La Jolla, USA
  • Year and Date: 2014-08-04 – 2014-08-05
  • Invited
• [Presentation] DNA脱メチル化に伴うDNA損傷機構へのヘミメチル化DNA結合タンパク質UHRF1の関与 (2014)
  • Author(s): 柴田隆豊、吉村健、栗谷健志、緒方進、奥村克純
  • Organizer: 日本生化学会中部支部第77回例会
  • Place of Presentation: 名古屋大学野依記念学術交流会館(名古屋市千種区)
  • Year and Date: 2014-05-24
• [Presentation] DNA低メチル化に伴うDNA損傷誘導機構について －DNAメチル化阻害剤5-aza-2’-deoxycytidine処理によるDNMT1の減少－ (2014)
  • Author(s): 村林雄太、金岡秀明、栗谷健志、緒方進、奥村克純
  • Organizer: 日本生化学会中部支部第77回例会
  • Place of Presentation: 名古屋大学野依記念学術交流会館(名古屋市千種区)
  • Year and Date: 2014-05-24
• [Presentation] 複製依存的DNA脱メチル化後のDNA損傷誘導機構 -hairpin-bisulfite sequence法によるヘミメチル化レベル解析- (2014)
  • Author(s): 生野彰宏、吉村健、緒方進、奥村克純
  • Organizer: 日本生化学会中部支部第77回例会
  • Place of Presentation: 名古屋大学野依記念学術交流会館(名古屋市千種区)
  • Year and Date: 2014-05-24
• [Presentation] 受動的DNA脱メチル化に伴うDNA損傷誘導機構の解明 (2014)
  • Author(s): 奥村 克純、栗谷 健志、生野 彰宏、吉村 健、来馬 啓介、伊藤 克
  • Organizer: 日本農芸化学会2014年度大会
  • Place of Presentation: 明治大学生田キャンパス
• [Presentation] 複製依存的DNA脱メチル化後のDNA損傷機構―hairpin-bisulfite sequence法によるヘミメチル化レベル解析― (2013)
  • Author(s): 生野彰宏，吉村健，奥村克純
  • Organizer: 日本生化学会中部支部例会
  • Place of Presentation: 名古屋大学
• [Presentation] DNMT1 減少時における DNA 損傷及び複製フォーク停止機構 (2013)
  • Author(s): 栗谷健志、来馬啓介、吉村健、奥村克純
  • Organizer: 第31回染色体ワークショップ・第12回核ダイナミクス研究会
  • Place of Presentation: 足柄下郡箱根町