Development of Optochemical Biology for photo-mediated regulation of gene function

• Project/Area Number: 26282219
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Basic / Social brain science
• Research Institution: Toho University
• Principal Investigator: FURUTA Toshiaki 東邦大学, 理学部, 教授 (90231571)
• Research Collaborator: SUZUKI Akinobu
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥16,640,000 (Direct Cost: ¥12,800,000、Indirect Cost: ¥3,840,000); Fiscal Year 2016: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2015: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2014: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
• Keywords: ケージド化合物 / 光化学 / 遺伝子 / 有機合成化学 / DNA / RNA / 遺伝子発現 / 有機化学 / ゲノム編集 / クリック反応 / PCR

Outline of Final Research Achievements

Photochemical activation of caged oligonucleotides enable the regulation of genes of interest within a single-cell level of spatial resolution. In the course of our study of the development of caged oligonucleotides, we designed and synthesized new precursor molecules of photo-releasable nucleotides having an affinity tag for purification and detection. A precursor molecule, Bio-Bhc-diazo comprises three components: biotin for an affinity tag, photoactive Bhc group, and phosphate reactive diazomethyl moiety. We developed a method to modify arbitrary sequence regions of long-chain double-stranded DNA and plasmid DNA with photolabile protecting groups by chemo-enzymatic reactions. We demonstrated that a caged compound of RNA can be synthesized using a novel nucleotide caging reagent having a biotin tag. We demonstrated that this technique can be applied to the gRNA of CRISPR / Cas9 system to synthesize caged gRNA protecting target DNA recognition site or Cas9 binding site.

Research Products

• [Journal Article] A double bond-conjugated dimethylnitrobenzene-type photolabile nitric oxide donor with improved two-photon cross section (2015)
  • Author(s): 1.N. Ieda, K. Hishikawa, K. Eto, K. Kitamura, M. Kawaguchi, T. Suzuki, K. Fukuhara, N. Miyata, T. Furuta, J. Nabekura, H. Nakagawa
  • Journal Title: Bioorg. Med. Chem. Lett. Volume: 25 Issue: 16 Pages: 3172-3175
  • DOI: 10.1016/j.bmcl.2015.05.095
  • Peer Reviewed / Open Access
• [Journal Article] Utilization of the Heavy Atom Effect for the Development of a Photosensitive 8-Azacoumarin-Type Photolabile Protecting Group (2015)
  • Author(s): Hikaru Takano, Tetsuo Narumi, Wataru Nomura, Toshiaki Furuta, and Hirokazu Tamamura
  • Journal Title: Organic Letters Volume: 17 Issue: 21 Pages: 5372-5375
  • DOI: 10.1021/acs.orglett.5b02720
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Synthesis of nucleobase-caged peptide nucleic acids having improved photochemical properties (2014)
  • Author(s): T. Watanabe, T. Hoshida, J. Sakyo, M. Kishi, S. Tanabe, J. Matsuura, S. Akiyama, M. Nakata, Y. Tanabe, A. Suzuki, S. Watanabe, T. Furuta
  • Journal Title: Org. Biomol. Chem. Volume: 12 Issue: 28 Pages: 5089-5093
  • DOI: 10.1039/c4ob00418c
  • Peer Reviewed
• [Journal Article] Development of the 8-aza-3-bromo-7-hydroxycoumarin-4-ylmethyl group as a new entry of photolabile protecting groups (2014)
  • Author(s): Takano, H., Narumi, T., Ohashi, N., Suzuki, A., Furuta, T., Nomura, W., Tamamura, H.
  • Journal Title: Tetrahedron Volume: XX Issue: 29 Pages: 4400-4404
  • DOI: 10.1016/j.tet.2014.04.063
  • Peer Reviewed
• [Presentation] Design and synthesis of caged compounds with cell type specificity (2017)
  • Author(s): T. Furuta, A. Z. Suzuki, T. Sakano
  • Organizer: 日本薬学会第137年会
  • Place of Presentation: 仙台国際会議場（宮城県仙台市）
  • Year and Date: 2017-03-27
  • Invited
• [Presentation] Caged compounds as optochemical genetic tools －Design, synthesis and their use－ (2016)
  • Author(s): T. Furuta
  • Organizer: The 17th RIES-HOKUDAI International Symposium on Ju
  • Place of Presentation: シャトレーゼガトーキングダムサッポロ（北海道札幌市）
  • Year and Date: 2016-12-14
  • Int'l Joint Research / Invited
• [Presentation] ynthesis of caged gRNAs for photoactivatable CRISPR-Cas9 (2016)
  • Author(s): K. Kodama, S. Tada, A. Z. Suzuki, T. Furuta
  • Organizer: The Fourth Asian Chemical Biology Conference (ACBC2016)
  • Place of Presentation: 85 Sky Tower Hotel (Kaohsiung, TAIWAN)
  • Year and Date: 2016-11-30
  • Int'l Joint Research
• [Presentation] Design and synthesis of site-selectively modified caged plasmid DNAs using modular nucleotide caging agents (2016)
  • Author(s): S. Tada, A. Z. Suzuki, T. Furuta
  • Organizer: The Fourth Asian Chemical Biology Conference (ACBC2016)
  • Place of Presentation: 85 Sky Tower Hotel (Kaohsiung, TAIWAN)
  • Year and Date: 2016-11-30
  • Int'l Joint Research
• [Presentation] CRISPR/Cas9の光制御を目指したケージドgRNAの合成 (2016)
  • Author(s): 児玉一徳・多田慎之介・鈴木商信・古田寿昭
  • Organizer: 第6回CSJ化学フェスタ2016
  • Place of Presentation: タワーホール船堀（東京都江戸川区）
  • Year and Date: 2016-11-16
• [Presentation] Synthesis of caged gRNA for photoactivatable CRISPR/Cas9 (2016)
  • Author(s): K. Kodama, S. Tada, A. Suzuki, T. Furuta
  • Organizer: 日本化学会 第96春季年会 (2016)
  • Place of Presentation: 同志社大学京田辺キャンパス（京都府・京田辺市）
  • Year and Date: 2016-03-26
• [Presentation] Design and synthesis of caged oligonucleotides with modular nucleotide caging agents (2016)
  • Author(s): S. Tada, A. Suzuki, T. Furuta
  • Organizer: 日本化学会 第96春季年会 (2016)
  • Place of Presentation: 同志社大学京田辺キャンパス（京都府・京田辺市）
  • Year and Date: 2016-03-26
• [Presentation] Modular caging groups – new platforms for multifunctional caged compounds (2015)
  • Author(s): T. Furuta
  • Organizer: The International Chemical Congress of Pacific Basin Societies 2015 (Pacifichem 2015)
  • Place of Presentation: Hawaii (USA)
  • Year and Date: 2015-12-20
  • Int'l Joint Research / Invited
• [Presentation] Chemo-enzymatic preparation of site-selectively modified caged DNAs (2015)
  • Author(s): S. Tada, W. Hashiba, E. Ikeda, A. Suzuki, T. Furuta
  • Organizer: The International Chemical Congress of Pacific Basin Societies 2015 (Pacifichem 2015)
  • Place of Presentation: Hawaii (USA)
  • Year and Date: 2015-12-19
  • Int'l Joint Research
• [Presentation] Design and synthesis of clickable caged compounds (2015)
  • Author(s): A. Suzuki, R. Aikawa, K. Kakehi, T. Furuta
  • Organizer: The International Chemical Congress of Pacific Basin Societies 2015 (Pacifichem 2015)
  • Place of Presentation: Hawaii (USA)
  • Year and Date: 2015-12-19
  • Int'l Joint Research
• [Presentation] プラスミドDNAに領域選択的に光分解性保護基を導入する手法の開発 (2015)
  • Author(s): 多田慎之介，古田寿昭
  • Organizer: 第5回CSJ化学フェスタ（2015）
  • Place of Presentation: タワーホール船堀（東京都・江戸川区）
  • Year and Date: 2015-10-15
• [Presentation] Design and synthesis of new caged diacylglycerol (2015)
  • Author(s): 池谷柚季，森有里紗，鈴木商信，古田 寿昭
  • Organizer: 日本化学会第95春季年会(2015)
  • Place of Presentation: 日本大学理工学部船橋キャンパス／薬学部（千葉県船橋市）
  • Year and Date: 2015-03-27
• [Presentation] Chemical tools to control cellular chemistry (2015)
  • Author(s): Toshiaki Furuta
  • Organizer: 第92回 日本生理学会大会
  • Place of Presentation: 神戸国際会議場・展示場（兵庫県神戸市）
  • Year and Date: 2015-03-22
  • Invited
• [Presentation] ケージドDNAを化学合成するためのホスホロアミダイト試薬の合成 (2014)
  • Author(s): 池田英一郎，鈴木商信，古田寿昭
  • Organizer: 第4回CSJ化学フェスタ
  • Place of Presentation: タワーホール船堀（東京都江戸川区）
  • Year and Date: 2014-10-16
• [Remarks] 光で細胞機能を操作するOptochemical Biologyの創生
  • URL: http://www.lab.toho-u.ac.jp/sci/biomol/tfuruta-lab/