Structural probing of RNA molecules with in-cell NMR
| Project/Area Number |
24310163
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Partial Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Chemical biology
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| Research Institution | Tokushima Bunri University (2015)
Tohoku University (2012-2014) |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
KOJIMA Chojiro 横浜国立大学, 大学院工学研究科, 教授 (50333563)
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| Co-Investigator(Renkei-kenkyūsha) |
TOCHIO Hidehito 京都大学, 大学院理学研究科, 教授 (70336593)
ONO Akira 神奈川大学, 工学部, 教授 (10183253)
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| Project Period (FY) |
2012-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥19,500,000 (Direct Cost: ¥15,000,000、Indirect Cost: ¥4,500,000)
Fiscal Year 2014: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2013: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
Fiscal Year 2012: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000)
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| Keywords | 生体分子計測 / NMR / 安定同位体標識 / バイオプローブ / 細胞内化学反応 |
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| Outline of Final Research Achievements |
In-cell NMR has been rarely applied to nucleic acids. In order to apply in-cell NMR into nucleic acids, there are several issues. 1) There has been no appropriate site-specific labeling technique of long RNA molecules. 2) There was no established method for introducing nucleic acids into cells. Therefore, at first, Tanaka (principal investigator: PI) developed a site-specific labeling technique of RNA molecules by using a recently discovered primer-extension ability of RNA polymerases. Interestingly, by using the derived labeled RNA molecule (HAC1 mRNA), PI identified that conserved C883 and G888 residues were base-paired within a loop. Then, in collaboration with grant team members, a method for introducing RNA molecules into cells was established by using an electroporation technique. It should be also mentioned that chemical reactions occurred to the introduced molecule was detected by using appropriate 19F-labeling tag. Thus, the team accomplished main purposes of this project.
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Report
(5 results)
Research Products
(48 results)
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[Journal Article] Direct Detection of the Mercury-Nitrogen Bond in the Thymine-HgII-Thymine Base-pair with 199Hg NMR Spectroscopy2015
Author(s)
Takenori Dairaku, Kyoko Furuita, Hajime Sato, Jakub Sebera, Daichi Yamanaka, Hiroyuki Otaki, Shoko Kikkawa, Yoshinori Kondo, Ritsuko Katahira, F. Matthias Bickelhaupt, Celia Fonseca Guerra, Akira Ono, Vladimir Sychrovsky, Chojiro Kojima and Yoshiyuki Tanaka
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Journal Title
Chemical Communications
Volume: 51
Issue: 40
Pages: 8488-8491
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] Structures, physicochemical properties, and applications of T-HgII-T, C-AgI-C, and other metallo-base-pairs.2015
Author(s)
Yoshiyuki Tanaka, Jiro Kondo, Vladimir Sychrovsky, Jakub Sebera, Takenori Dairaku, Hisao Saneyoshi, Hidehito Urata, Hidetaka Torigoe and Akira Ono
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Journal Title
Chem. Commun.
Volume: 51
Issue: 98
Pages: 17313-17360
DOI
Related Report
Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
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[Journal Article] NMR spectroscopic studies on metallo-base-pair in DNA duplex.2014
Author(s)
Yoshiyuki Tanaka, Itaru Okamoto, Kyoko Furuita, Jakub Sebera, Jiro Kondo, Hidetaka Torigoe, Hidehito Urata, Takenori Dairaku, Akira Ono, Chojiro Kojima, Vladimir Sychrovsky
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Journal Title
Journal of Biological Inorganic Chemistry
Volume: 19
Issue: S2
Pages: 725-747
DOI
Related Report
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[Journal Article] Proton and nitrogen-15 NMR spectroscopic studies of AgI-mediated C-C base-pairs.2014
Author(s)
Takenori Dairaku, Itaru Okamoto, Kyoko Furuita, Shuji Oda, Daichi Yamanaka, Hidetaka Torigoe, Tetsuo Kozasa, Yoshinori Kondo, Akira Ono, Chojiro Kojima, Vladimir Sychrovsky, Yoshiyuki Tanaka
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Journal Title
Journal of Biological Inorganic Chemistry
Volume: 19
Issue: S2
Pages: 815-832
DOI
Related Report
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[Journal Article] Raman spectroscopic detection of the T-HgII-T base pair and the ionic characteristics of mercury2012
Author(s)
Tomomi Uchiyama, Takashi Miura, Hideo Takeuchi, Takenori Dairaku, Tomoyuki Komuro, Takuya Kawamura, Yoshinori Kondo, Ladislav Benda, Vladimir Sychrovsky, Petr Bour, Itaru Okamoto, Akira Ono and Yoshiyuki Tanaka
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Journal Title
Nucleic Acids Res
Volume: 40
Issue: 12
Pages: 5766-5774
DOI
Related Report
Peer Reviewed
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[Presentation] NMR spectroscopic studies on metallo-base-pair in DNA duplex.2014
Author(s)
◯Yoshiyuki Tanaka, Itaru Okamoto, Kyoko Furuita, Jakub Sebera, Jiro Kondo, Hidetaka Torigoe, Hidehito Urata, Takenori Dairaku, Akira Ono, Chojiro Kojima, Vladimir Sychrovsky
Organizer
12th European Biological Inorganic Chemistry Conference (EuroBIC-12)
Place of Presentation
チューリッヒ、スイス
Year and Date
2014-08-24 – 2014-08-28
Related Report
Invited
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[Presentation] Proton and nitrogen-15 NMR spectroscopic studies of AgI-mediated C-C base-pairs.2014
Author(s)
◯Takenori Dairaku, Itaru Okamoto, Kyoko Furuita, Shuji Oda, Daichi Yamanaka, Hidetaka Torigoe, Tetsuo Kozasa, Yoshinori Kondo, Akira Ono, Chojiro Kojima, Vladimir Sychrovsky, Yoshiyuki Tanaka
Organizer
12th European Biological Inorganic Chemistry Conference (EuroBIC-12)
Place of Presentation
チューリッヒ、スイス
Year and Date
2014-08-24 – 2014-08-28
Related Report
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[Presentation] 水銀イオンのT-Tミスマッチ塩基対に対する親和性の構造化学的要因（口頭発表）2013
Author(s)
◯田中好幸, 大樂武範, 根東義則, 内山朋美, 三浦隆史, 竹内英夫, 古板恭子, 児嶋長次郎, Ladislav Benda, Jakub Sebera, Vladimir Sychrovsky, 岡本到, 小野晶
Organizer
第回 生体分子科学討論会
Place of Presentation
大阪
Related Report
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[Presentation] Structure of Ag(I)-mediated C-C base pair determined by hetero-nuclear NMR spectroscopy2013
Author(s)
Takenori Dairaku, Itaru Okamoto, Kyoko Furuita, Jukub Sebera, Mathias Bickelhaupt, Shuji Oda, Daichi Yamanaka, Yoshinori Kondo, Vladimir Sychrovsky, Chojiro Kojima, Akira Ono, ◯Yoshiyuki Tanaka
Organizer
40th International Symposium on Nucleic Acids Chemistry
Place of Presentation
横浜
Related Report
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