Chemical splicing for regulation of functional nucleic acids and cells

• Project/Area Number: 15K13749
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Bio-related chemistry
• Research Institution: Kumamoto University
• Principal Investigator: Ihara Toshihiro 熊本大学, 大学院先端科学研究部(工), 教授 (40253489)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: DNA conjugate / terpyridine / anthracene / sequence edition / metal ion / photo dimerization / omega-shaped structure / chemical splicing / アントラセン / 光二量化 / 化学的スプライシング / 金属イオン / 光 / DNAzyme / ターピリジン / DNAzyme活性制御 / DNAの構造制御 / Ω型DNA / DNAコンジュゲート / 化学的DNA編集

Outline of Final Research Achievements

Two terpyridine units were built into the distal sites of a DNA backbone to obtain the conjugates. The conjugate formed an intramolecular 1:2 complex with some transition metal ions such as Fe3+ and Ni3+ to take an Ω-shaped structure. The two sequences located outsides of two terpyridine units was connected directly with each other to provide a new sequence. That is, the continuity of distal segments was controlled by metal ion-induced Ω-shaped conformation. We succeeded in the activity control of a split DNAzyme using this techniques of metal ion-directed reversible sequence edition.
Two anthracene units were built into the DNA backbone to obtain the conjugates. The conjugate formed an Ω-shaped structure through intramolecular dimerization of anthracene by photoirradiation in the presence of the template DNA carrying the sequence complementary to flanking two sequences of anthracene units. This could be regarded as photo-switchable techniques of reversible sequence edition.

Research Products

• [Journal Article] Electrochemical Sensing of Neurotoxic Agents Based on Their Electron Transfer Promotion Effect on an Au Electrode (2017)
  • Author(s): Hiroshi Shimada, Shiori Noguchi, Masahiro Yamamoto, Katsuhiko Nishiyama, Yusuke Kitamura, and Toshihiro Ihara
  • Journal Title: Analytical Chemistry Volume: 89 Issue: 11 Pages: 5742-5747
  • DOI: 10.1021/acs.analchem.6b04229
  • Peer Reviewed
• [Journal Article] Sensitive Electrochemical Detection of Nereistoxin by Reductive Desorption from Au(111) and Au(100) (2016)
  • Author(s): K. Nishiyama, K. Tsuruta, M. Ikeda, S. Yoshimoto, H. Shimada, Y. Kitamura, T. Ihara
  • Journal Title: Electrochemistry Volume: 84 Issue: 5 Pages: 349-353
  • DOI: 10.5796/electrochemistry.84.349
  • NAID: 130005150422
  • ISSN: 1344-3542, 2186-2451
  • Peer Reviewed / Open Access
• [Journal Article] Metal Ion-directed Dynamic Splicing of DNA through Global Conformational Change by Intramolecular Complexation (2015)
  • Author(s): T. Ihara, H. Ohura, C. Shirahama, T. Furuzono, H. Shimada, H. Matsuura, Y. Kitamura
  • Journal Title: Nature Communications Volume: 6 Issue: 1 Pages: 6640-6640
  • DOI: 10.1038/ncomms7640
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Graphene Oxide-based Amplified Fluorescence Sensor for Nucleic Acid Detection through Target-catalyzed Hairpin Assembly (2015)
  • Author(s): Y. Kitamura, T. Miyahata, H. Matsuura, K. Hatakeyama, T. Taniguchi, M. Koinuma, Y. Matsumoto, T. Ihara
  • Journal Title: Chemistry Letters Volume: 44 Issue: 10 Pages: 1353-1355
  • DOI: 10.1246/cl.150564
  • NAID: 130005101443
  • Peer Reviewed
• [Presentation] 外部刺激によるコンフォメーション変化を利用したDNA塩基配列の可逆的編集 (2017)
  • Author(s): 大塚幸貴広、大浦博之、北村祐介、井原敏博
  • Organizer: 日本化学会第97春季年会2017
  • Place of Presentation: 慶應大学
  • Year and Date: 2017-03-16
• [Presentation] 核酸を基体とするバイオセンシング (2017)
  • Author(s): 井原敏博
  • Organizer: 高分子学会九州支部 有機材料研究グループ研究会
  • Invited
• [Presentation] Target Recognition by Global DNA Structural Control (2017)
  • Author(s): Toshihiro Ihara
  • Organizer: FIBER International Summit for Nucleic Acids 2017
  • Int'l Joint Research / Invited
• [Presentation] 人工DNAの構造制御 バイオ分析・機能制御を目指して (2017)
  • Author(s): 井原敏博
  • Organizer: 大阪府立大学大学院理学系研究科講演会
  • Invited
• [Presentation] Capture of cancer cells on a gold substrate modified with DNA aptamer (2017)
  • Author(s): Y. Kitamura, A. Tashima, M. Nakamura, Y. Nakashima, K. Yasuda, M. Iwatsuki, Y. Katsuda, H. Baba, Y. Nakanishi, T. Ihara
  • Organizer: International Symposium on Nucleic Acids Chemistry 2017
  • Int'l Joint Research
• [Presentation] 核酸を基体とするバイオセンシン グ (2017)
  • Author(s): 井原敏博
  • Organizer: 高分子学会九州支部 有機材料研究グループ研究会
  • Place of Presentation: 九州大学
  • Invited
• [Presentation] DNA の動的プログラミングを利用したバイオセンシング (2016)
  • Author(s): 井原敏博
  • Organizer: 日本分析化学会第65年会
  • Place of Presentation: 北海道大学
  • Year and Date: 2016-09-14
  • Invited
• [Presentation] Metal Complexation on DNA -For DNA Structural Control and Biosensing- (2016)
  • Author(s): Toshihiro Ihara
  • Organizer: FIBER International Summit for Nucleic Acids 2016
  • Place of Presentation: 甲南大学
  • Year and Date: 2016-07-06
  • Int'l Joint Research / Invited
• [Presentation] BIOSENSING BASED ON RARE EARTH METAL COMPLEX FORMATION THROUGH DYNAMIC PROGRAMMING OF DNA STRUCTURE (2016)
  • Author(s): Toshihiro Ihara, Yukina Azuma, Rie Ozaki, Akihiro Nozaki, Takaaki Miyahata, Yusuke Kitamura
  • Organizer: Rare Earths 2016 in Sapporo
  • Place of Presentation: 北海道大学
  • Year and Date: 2016-06-05
  • Int'l Joint Research / Invited
• [Presentation] Cooperative formation of metal complexes on nucleic acids (2016)
  • Author(s): Yusuke Kitamura, Yukina Azuma, Shikinari Yamamoto,Toshihiro Ihara
  • Organizer: 7th International Symposium on Bioanalysis, Biomedical Engineering and Nanotechnology
  • Place of Presentation: Hunan University
  • Year and Date: 2016-05-27
  • Int'l Joint Research / Invited
• [Presentation] DNA上での錯生成反応 -バイオ分析・機能制御を目指して- (2016)
  • Author(s): 井原敏博
  • Organizer: 名古屋大学大学院物質制御工学選考セミナー
  • Place of Presentation: 名古屋大学
  • Invited
• [Presentation] 核酸上での錯生成反応 –核酸の構造制御およびバイオ分析への応用– (2015)
  • Author(s): 井原敏博
  • Organizer: 和光ライフサイエンスフォーラム
  • Place of Presentation: 熊本市
  • Year and Date: 2015-12-11
  • Invited
• [Presentation] METAL COMPLEXATION ON DNA FOR DNA STRUCTURE CONTROL AND BIOSENSING (2015)
  • Author(s): Toshihiro Ihara
  • Organizer: 16th IUPAC International Symposium on MacroMolecular Complexes (MMC-16)
  • Place of Presentation: Wroclaw
  • Year and Date: 2015-08-10
  • Int'l Joint Research / Invited
• [Presentation] 核酸の構造制御およびバイオセンシングへの応用 (2015)
  • Author(s): 井原敏博
  • Organizer: 核酸化学最前線フォーラム
  • Place of Presentation: 神戸市
  • Year and Date: 2015-07-09
  • Invited
• [Presentation] DNA構造の動的プログラミングに基づくバイオセンシング (2015)
  • Author(s): 井原敏博
  • Organizer: 第52回化学関連支部合同九州大会
  • Place of Presentation: 北九州市
  • Year and Date: 2015-06-27
  • Invited
• [Book] Advances in Bioorganometallic Chemistry (2019)
  • Author(s): Toshihiro Ihara
  • Total Pages: 300
  • Publisher: Elsevier
• [Remarks] 熊本大学井原研究室ウェブサイト
  • URL: http://www.analyticalchemistry-ihara.com
• [Remarks] IHARA Lab@Kumamoto U
  • URL: http://133.95.131.186/~toshi/
• [Remarks] IHARA Lab@Kumamoto-U
  • URL: http://133.95.131.186/~toshi/