Design of Base-discriminating Fluorescent Nucleosides and Their Application to Gene Analysis and SNP Genotyping

• Project/Area Number: 18201026
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Nanomaterials/Nanobioscience
• Research Institution: Nihon University
• Principal Investigator: SAITO Isao Nihon University, 工学部, 教授 (20026082)
• Co-Investigator(Kenkyū-buntansha): SAITO Yoshio 日本大学, 工学部, 准教授 (40385985)
• Project Period (FY): 2006 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥47,970,000 (Direct Cost: ¥36,900,000、Indirect Cost: ¥11,070,000); Fiscal Year 2009: ¥7,150,000 (Direct Cost: ¥5,500,000、Indirect Cost: ¥1,650,000); Fiscal Year 2008: ¥10,140,000 (Direct Cost: ¥7,800,000、Indirect Cost: ¥2,340,000); Fiscal Year 2007: ¥12,220,000 (Direct Cost: ¥9,400,000、Indirect Cost: ¥2,820,000); Fiscal Year 2006: ¥18,460,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥4,260,000)
• Keywords: DNAデバイス / DNAチップ / 遺伝子検出 / 遺伝子診断 / SNP検出 / ナノバイオ / ゲノム / 蛍光分子

Research Abstract

We have designed novel base-discriminating fluorescent (BDF) nucleosides which are successfully used for SNP genotyping and gene detection. Quencher and end free molecular beacons (MB) were discovered for the first time. These low cost MB were used for the gene detection in a cell. We have also succeeded to design environmentally sensitive fluorescent nucleosides that emits strong fluorescence at different wavelengths depending upon a local environment around the nucleobase.

Research Products

• [Journal Article] Fluorescence Switching of Photochromic Vinylpyrene Substituted 2'-Deoxyguanosine. (2009)
  • Author(s): Y. Saito, K. Matsumoto, Y. Takeuchi, S.S. Bag, S. Kodate, T. Morii, I. Saito
  • Journal Title: Tetrahedron Lett. 50 Pages: 1403-1406
  • Peer Reviewed
• [Journal Article] Ends Free and Self-quenched Molecular Beacons with Pyrene Labeled Pyrrolocitidine in the Middle of the Stem. (2009)
  • Author(s): Y. Saito, Y. Sinohara, S.S. Bag, Y. Takeuchi, K. Matsumoto, I. Saito
  • Journal Title: Terahedron 65 Pages: 934-934
  • Peer Reviewed
• [Journal Article] Ends Free and Self-quenched Molecular Beacons with Pyrene Labeled Pyrrolocitidine in the Middle of the Stem (2009)
  • Author(s): Y.Saito, Y.Sinohara, S.S.Bag, Y.Takeuchi, K.Matsumoto, I.Saito
  • Journal Title: Terahedron 65 Pages: 934-938
  • Peer Reviewed
• [Journal Article] Fluorescence Switching of Photochromic Vinylpyrene Substituted 2'-Deoxyguanosine (2009)
  • Author(s): Y.Saito, K.Matsumoto, Y.Takeuchi, S.S.Bag, S.Kodate, T.Morn, I.Saito
  • Journal Title: Tetrahedron Lett 50 Pages: 1403-1406
  • Peer Reviewed
• [Journal Article] Design of Extremely Facile 3'- and 5'- Ends Free Molecular Beacons using C8 Alkylamino Substituted 2'-Deoxyguanosine (2009)
  • Author(s): K.Matsumoto, Y.Shinohara, K.Numajiri, S.Ishioroshi, T.Morii, Y.Saito, I.Saito
  • Journal Title: Nucleic Acids Symp Ser 53 Pages: 141-142
• [Journal Article] C8-Alkynyl-and Alkylamino Substituted 2'-Deoxyguanosines: A Universal Linker for Nucleic Acids Modification (2008)
  • Author(s): Y. Saito, K. Matsumoto, S.S. Bag, S. Ogasawara, K. Fujimoto, K. Hanawa, I. Saito
  • Journal Title: Tetrahedron 64 Pages: 3578-3488
  • Peer Reviewed
• [Journal Article] Anthracene Based Base-discriminating Fluorescent Oligonucleotide Probes for SNPs Typing: Synthesis and Photophysical Properties (2008)
  • Author(s): Y. Saito, K. Motegi, S.S. Bag, I. Saito
  • Journal Title: Bioorg. Med. Chem. Symposium-in-Print 16 Pages: 107-113
  • Peer Reviewed
• [Journal Article] C8-Alkynyl and alkylamino substituted 2'-deoxyguanosine (2008)
  • Author(s): Y. Saito, K. Matsumoto, S. S. Bag, K. Hanawa and I. Saito
  • Journal Title: Tetrahedron 64 Pages: 3478-3488
• [Journal Article] Anthracene based base-discriminating fluorescence ologonucleotide probe SNP (2008)
  • Author(s): Y, Saito, K. Motegi, S.S. Bag, and I. Saito
  • Journal Title: Biooorg. Med. Chem. 16 Pages: 107-113
• [Journal Article] Sysnthesis and reversible photoisomerization of photoswitchable nucleoside 8-styryll-2'-deoxyguanosine (2008)
  • Author(s): S. Ogasawara, I. Saito, and M. Maeda
  • Journal Title: Tetrahedron Lett. 49 Pages: 2479-2482
• [Journal Article] Anthracene Based Base-discriminating Fluorescent Oligonucleotide Probes for SNPs Typing : Synthesis and Photophysical Properties (2008)
  • Author(s): Y. Saito, K. Motegi, S. S. Bag, I. Saito.
  • Journal Title: Bioorg. Med. Chem. Syaposium-in-Print 16 Pages: 107-113
  • Peer Reviewed
• [Journal Article] Synthesis and Reversible Photoisomerization of Photoswitchable Nucleoside 8-Styryl-2'- deoxyguanosine (2008)
  • Author(s): S. Ogasawa, I. Saito, M. Maeda
  • Journal Title: Tetrahedron Lett. 49 Pages: 2479-2482
  • Peer Reviewed
• [Journal Article] C8-Alkynyl- and Alkylamino Substituted 2'-Deoxyguanosines : A Universal Linker for Nucleic Acids Modification (2008)
  • Author(s): Y. Saito, K. Matsumoto, S. S Bag, S. Ogasawara, K. Fujimoto, K. Hanawa, I. Saito
  • Journal Title: Tetrahedron 64 Pages: 3578-3488
  • Peer Reviewed
• [Journal Article] Design of a Novel G-quenched Molecular Beacon; A Simple and Efficient Strategy for DNA Sequenceing Analysis (2007)
  • Author(s): Y. Saito, E. Mizuno, S.S. Bag, I. Suzuka, I. Saito.
  • Journal Title: Chem. Commun. Pages: 4492-4494
  • Peer Reviewed
• [Journal Article] Reversible Photopadlocking on Double-stranded DNA. (2007)
  • Author(s): K. Fujimoto, S. Matsuda, Y. Yoshimura, T. Ami, I. Saito
  • Journal Title: Chem. Commun. Pages: 2968-2970
  • Peer Reviewed
• [Journal Article] PRODAN-Conjugated DNA: Synthesis and Photochemical Properties (2007)
  • Author(s): K. Tainaka, K. Tanaka, Ikeda, K-i., Nishiza, T. Unzai, Y. Fujiwara, I. Saito, A. Okamoto
  • Journal Title: J. Am. Chem. Soc 129 Pages: 4776-4784
  • Peer Reviewed
• [Journal Article] Synthesis and Fluorescence Properties of Dimethylaminonaphthalene-deoxyuridine Conjugates As Micropolarity-sensitive Probes. (2007)
  • Author(s): Okamoto, A. ; Tainaka, K. ; Unzai, T. ; Saito
  • Journal Title: I. Tetrahedron 63 Pages: 3465-3470
  • Peer Reviewed
• [Journal Article] Dual-labeled Oligonucleotide Probe for Sensing Adenosine via FRET (2007)
  • Author(s): Y. Saito, S. S. Bag, Y. Kusakabe, C. Nagai, K. Matsumoto, S. Kodate, I. Suzuka and I. Saito
  • Journal Title: Chem. Commun. Pages: 2133-2135
  • Peer Reviewed
• [Journal Article] Anthracene Based Base-discriminating Fluorescent Oligonucleotide Probes for SNPs Typing (2007)
  • Author(s): Y. Saito, K. Motegi, S. S. Bag, and I. Saito
  • Journal Title: Bioorg. Med. Chem. Symposisum-in-Print 16 Pages: 107-113
  • Peer Reviewed
• [Journal Article] Design of a Novel G-quenched Molecular Beacons; A Simple and Efficient Strategy for DNA Sequencing Analysis (2007)
  • Author(s): Y. Saito, E. Mizuno, S. S. Bag, I. Suzuka and I. Saito
  • Journal Title: Chem. Commun. Pages: 4492-4493
  • Peer Reviewed
• [Journal Article] Highly Selective Fluorescent Nucleobases for Designing Base-Discriminating Fluorescence Probe. (2006)
  • Author(s): Saito, I. ; Saito, Y. ; Hanawa, K. ; Hayashi, K. ; Motegi, K. ; Bag, S.S. ; Dohno, C. ; Ichiba, T. ; Tainaka, K. ; Okamoto
  • Journal Title: A. Pure. Appl. Chemistry 78(12) Pages: 2305-2312
  • Peer Reviewed
• [Journal Article] Simple SNP Typing Assay Using a Base-discriminating Fluorescent Probe. (2006)
  • Author(s): A. Okamoto, K. Tainaka, Y. Ochi, K. Kanatani, I. Saito
  • Journal Title: Molecular BioSystem. 2 Pages: 81-81
  • Peer Reviewed
• [Journal Article] DNA Hole Transport on an Electrode : Application to Effective photoelectrochemical SNP Typing. (2006)
  • Author(s): A.Okamoto, T.Kamei, I.Saito
  • Journal Title: J. Am.Chem. Soc 128 Pages: 658-662
• [Journal Article] Acridone?labeled Base?discriminating Fluorescence Nuceloside (2006)
  • Author(s): Y.Saito, K.Hanawa, N.Kawasaki, S.S.Bag, I.Saito
  • Journal Title: Chemistry Letters 35 Pages: 1182-1183
• [Journal Article] Intelligent Fluorescent Nucleoside in Sensing Cytosine Base (2006)
  • Author(s): S.S.Bag, Y.Saito, K.Hanawa, S.Kodate, I.Suzuka, I.SAito
  • Journal Title: Bioorganic Med. Chem. Lett. 16 Pages: 6338-6342
• [Journal Article] Site?specific Transition of Cytosine to mUracil via Reversible DNA Photoligation (2006)
  • Author(s): K.Fujimoto, S.Matsuda, Y.Yoshimura, T.Matsumura, M.Hayashi, I.Saito
  • Journal Title: Chem. Commun Pages: 3223-3225
• [Presentation] 両末端フリーの自己消光型モレキュラービーコンの開発 (2009)
  • Author(s): 齋藤義雄, 篠原雄太, 松本桂彦, 沼尻恭子, 齋藤烈
  • Organizer: 2009年光化学討論会
  • Place of Presentation: 桐生市市民文化会館
  • Year and Date: 2009-09-16
• [Presentation] DNA検出のためのアントラセン、ピレン骨格を有するTwinプローブの開発 (2009)
  • Author(s): 齋藤義雄, 鈴木梓, 篠原雄太, 石下真也, 今井和俊, 根本修克, 齋藤烈
  • Organizer: 2009年光化学討論会
  • Place of Presentation: 桐生市市民文化会館
  • Year and Date: 2009-09-16
• [Presentation] 新しいSolvatofluorochromicなプリン核酸塩基のデザインと応用 (2009)
  • Author(s): 齋藤義雄, 高橋尚也, 松本桂彦, 篠原雄太, 竹内辰樹, 齋藤烈
  • Organizer: 2009年光化学討論会
  • Place of Presentation: 桐生市市民文化会館
  • Year and Date: 2009-09-16
• [Presentation] C8位にリンカーをもつ新規グアノシン誘導体の応用 (2009)
  • Author(s): 齋藤義雄, 松本桂彦, 篠原雄太, 沼尻恭子, 齋藤烈
  • Organizer: 日本化学会第89回春季年会
  • Place of Presentation: 日本大学理工学部船橋キャンパス
  • Year and Date: 2009-03-27
• [Presentation] ビニルピレン置換グアニン塩基を用いる蛍光スイッチの開発 (2009)
  • Author(s): 齋藤義雄, 竹内辰樹, 松本桂彦, 高橋尚也, 藤烈
  • Organizer: 日本化学会第89回春季年会
  • Place of Presentation: 日本大学理工学部船橋キャンパス
  • Year and Date: 2009-03-27
• [Presentation] Fluorometric sensing of the B-Z DNA transition of DNA Containing C8-aIkylainino substituted 2'-deoxyguanosine (2008)
  • Author(s): 松本 桂彦
  • Organizer: 第35回核酸化学国際シンポジウム
  • Place of Presentation: 京都大学吉田キャンパス
  • Year and Date: 2008-09-08
• [Presentation] Design of Dual-labeled Oligonucleotide Probes for SNPs Genotyping (2007)
  • Author(s): S. S. Bag
  • Organizer: 第5回国核酸シンポジュウム
  • Place of Presentation: 東大安田講堂
  • Year and Date: 2007-11-20
• [Presentation] アントラセン骨格を有するTwinプロープの開発と遺伝子検出への応用
  • Author(s): 篠原雄太、鈴木梓、石下真也、齋藤義雄、齋藤烈
• [Presentation] C8位にリンカーを持つ新規グアノシン誘導体の応用
  • Author(s): 松本桂彦、篠原雄太、沼尻恭子、齋藤義雄、齋藤烈、水野絵梨香、笠原竜、佐藤直樹、藤井美里
• [Presentation] ビニルピレン置換グアニン塩基を用いる蛍光スイッチの開発
  • Author(s): 竹内辰樹、松本桂彦、高橋尚也、齋藤義雄、齋藤烈
• [Book] 化学フロンティア18ゲノム化学の最先端 (2007)
  • Author(s): 齋藤烈, 杉山弘, 中谷和彦共編, 化学同人
• [Book] ゲノム化学の最先端 (2007)
  • Author(s): 齋藤烈、杉山弘、中谷和彦
  • Publisher: 化学同人
• [Remarks]
  • URL: http://ch.ce.nihon-u.ac.jp/%7ei_saito/EngTop.html
• [Remarks]
  • URL: http://ch.ce.nihon-u.ac.jp/%7ei_saito/EngTop.html
• [Remarks]
  • URL: http://ch.ce.nihon-u.ac.jp/%7ei_saito/EnTop.html
• [Remarks]
  • URL: http://ch.ce.nihon-u.ac.jp/%7ei_saito/EngToP.html
• [Patent(Industrial Property Rights)] c8位置換プリン塩基誘導体及びそれを利用した蛍光プローブ (2009)
  • Inventor(s): 齋藤烈, 齋藤義雄, 高橋尚弥
  • Industrial Property Rights Holder: 日本大学
  • Acquisition Date: 2009-08-17
• [Patent(Industrial Property Rights)] ステムループ構造を有する蛍光プローブ (2008)
  • Inventor(s): 齋藤烈, 齋藤義雄, 松本桂彦
  • Industrial Property Rights Holder: 日本大学
  • Acquisition Date: 2008-09-02
• [Patent(Industrial Property Rights)] ステムループ構造を有する蛍光プローブ (2008)
  • Inventor(s): 齋藤烈、齋藤義雄、松本桂彦
  • Industrial Property Rights Holder: 日本大学、長浜バイオラボラトリ(株)
  • Filing Date: 2008-09-02
• [Patent(Industrial Property Rights)] ステムループ構造を有する蛍光プロープ (2008)
  • Inventor(s): 齋藤烈齋藤義雄松本桂彦
  • Industrial Property Rights Holder: 日本大学
  • Filing Date: 2008-09-02
• [Patent(Industrial Property Rights)] ヌクレオチド誘導体及びその利用(FRET) (2007)
  • Inventor(s): 齋藤烈, 齋藤義雄, 吉田安子, 山田和成, 丹羽孝介
  • Industrial Property Rights Holder: 日本大学
  • Acquisition Date: 2007-02-08
• [Patent(Industrial Property Rights)] 水素結合性置換基を有する8―置換グアノシン誘導体及びそれを含むオリゴヌクレオチド (2006)
  • Inventor(s): 齋藤烈, 齋藤義雄, 花輪和夫
  • Industrial Property Rights Holder: 日本大学
  • Filing Date: 2006-12-27
• [Patent(Industrial Property Rights)] 光反応性核酸及び可逆的核酸光連結又は開裂方法 (2006)
  • Inventor(s): 齋藤烈, 齋藤義雄
  • Industrial Property Rights Holder: 日本大学
  • Acquisition Date: 2006-03-23