Development of tagged protein imaging during translation and folding steps by using genome editing
| Project/Area Number |
16K01931
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Chemical biology
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
Nomura Wataru 東京医科歯科大学, 生体材料工学研究所, 准教授 (80463909)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Keywords | ゲノム編集 / 蛍光イメージング / ペプチド / タンパク質 / タグ－プローブ / タグープローブ / 分子イメージング / ゲノム編集技術 |
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| Outline of Final Research Achievements |
In this project, a method to observe processes of protein translation, folding, and translocation was developed using our tag-probe system. Membrane receptor and histone protein were chosen as targets and their fusion with fluorescent protein were expressed in cells. Expression and localization of target proteins were observed by fluorescent microscopy. To introduce tag gene after starting codon, guide RNA was designed and genome editing by CRISPR-Cas9 was performed. We have established a method to select cells with drug-resistant marker to increase editing efficiency. The method can widely applied to introduce various gene to genome.
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| Academic Significance and Societal Importance of the Research Achievements |
細胞内に存在するタンパク質はその機能自体の解明も重要であるが，細胞の中にどれくらいの量が存在し，どこに存在するのか、存在場所は機能とともに変化するか，などを顕微鏡で観察することが必要になる。それを解決する手段として、細胞のゲノムに存在するタンパク質を定義している遺伝子に目印となるタグを付けることで細胞内のタンパク質の絶対量や動きを正確に観察することができる。本研究では独自のタグをゲノムに組み入れる方法を確立することができた。
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Report
(4 results)
Research Products
(49 results)
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[Journal Article] Benzolactam-related compounds promote apoptosis of HIV-infected human cells via protein kinase C-induced HIV latency reversal.2019
Author(s)
Kouki Matsuda, Takuya Kobayakawa, Kiyoto Tsuchiya, Shin-ichiro Hattori, Wataru Nomura, Hiroyuki Gatanaga, Kazuhisa Yoshimura, Shinichi Oka, Yasuyuki Endo, Hirokazu Tamamura, Hiroaki Mitsuya, Kenji Maeda
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Journal Title
Journal of Biological Chemistry
Volume: 294
Pages: 116-129
DOI
Related Report
Peer Reviewed
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[Journal Article] Design, synthesis and biological evaluation of low molecular weight CXCR4 ligands.2019
Author(s)
3.Sakyiamah MM., Kobayakawa T., Fujino M., KonnoM., Narumi T., Tanaka T., Nomura W., Yamamoto N., Murakami T., and Tamamura H.
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Journal Title
Bioorganic & Medicinal Chemistry
Volume: 2
Pages: 1130-1138
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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[Journal Article] A minimally cytotoxic CD4 mimic as an HIV entry inhibitor.2016
Author(s)
Takaaki Mizuguchi, Shigeyoshi Harada, Tomoyuki Miura, Nami Ohashi, Tetsuo Narumi, Hiromi Mori, Yu Irahara, Yuko Yamada, Wataru Nomura, Shuzo Matsushita, Kazuhisa Yoshimura, Hirokazu Tamamura
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Journal Title
Bioorg Med Chem Lett.
Volume: 26
Pages: 397-400
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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[Presentation] Protein dynamics imaging using tag-probe systems2016
Author(s)
Wataru Nomura, Nami Ohashi, HIrokazu Tamamura
Organizer
The 14th Chinese International Peptide Symposium & the 5th Asia-Pacific International Peptide Symposium
Place of Presentation
Nanjing, China
Year and Date
2016-07-06
Related Report
Int'l Joint Research / Invited
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