Identification and characterization of DNA/RNA heteroduplex oligonucleotide cleaving enzyme
| Project/Area Number |
18K05550
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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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| Review Section |
Basic Section 38060:Applied molecular and cellular biology-related
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| Research Institution | Institute of Physical and Chemical Research (2019-2020)
National Cancer Center Japan (2018) |
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Principal Investigator |
Asada Ken 国立研究開発法人理化学研究所, 革新知能統合研究センター, 研究員 (70773414)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | 核酸医薬 / 遺伝子発現制御 / ヌクレアーゼ / ヘテロ2本鎖核酸 / 一本鎖核酸 / 核酸医療 / DNA/RNAヘテロ核酸 (HDO) / ヘテロ核酸を用いた遺伝子制御 |
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| Outline of Final Research Achievements |
A tocopherol conjugated DNA/RNA heteroduplex oligonucleotide (Toc-HDO) as a new type of drug. Toc-HDO is more potent, stable, and efficiently taken up by the target tissues. However, the detailed mechanisms of Toc-HDO, including its binding proteins, are unknown. Here, we developed native gel shift assays with fluorescence-labeled nucleic acids samples extracted from mice livers. These assays revealed two Toc-HDO binding proteins, annexin A5 (ANXA5) and carbonic anhydrase 8 (CA8). Later, we identified two more proteins, apurinic/apyrimidinic endodeoxyribonuclease 1 (APEX1) and flap structure-specific endonuclease 1 (FEN1) by data mining. All four proteins regulated Toc-HDO activity in Hepa1-6, mouse hepatocellular cells. In vitro binding assays and pull-down assays demonstrated the protein binding to the Toc-HDO in a biological environment. Taken together, our findings provide a brand new molecular biological insight as well as future directions for HDO-based disease therapy.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究グループはこれまでに独自に開発した核酸医薬であるHDOが、非常に高い遺伝子発現抑制効果を示すことを報告しています。しかしながら、HDOの細胞内の作用分子メカニズムは解明されていませんでした。今回研究グループは、HDOに特異的な結合タンパク質を複数同定することに成功しました。同定したタンパク質は、塩基配列に関わらず13～16塩基対の短いDNA/RNAに選択的に結合できることを見出しました。同定した結合タンパク質が、HDOの活性制御に重要な働きをしていることを明らかにしました。
HDOの作用メカニズムの解明により、今後は効果が高く副作用の少ない核酸医薬の開発につながるものと期待されます。
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Report
(4 results)
Research Products
(20 results)
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[Journal Article] Two Secreted Proteoglycans, Activators of Urothelial Cell?Cell Adhesion, Negatively Contribute to Bladder Cancer Initiation and Progression2020
Author(s)
Papadaki V, Asada K, Watson JK, Tamura T, Leung A, Hopkins J, Dellett M, Sasai N, Davaapil H, Nik-Zainal S, Longbottom R, Nakakido M, Torii R, Veerakumarasivam A, Kaneko S, Sagoo MS, Murphy G, Mitani A, Tsumoto K, Kelly JD, Hamamoto R, Ohnuma SI
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Journal Title
Cancers
Volume: 12
Issue: 11
Pages: 3362-3362
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Presentation] Intefrative multi-omics analysis of multistage tumorigenesis in high-grade serous ovarian cancer2020
Author(s)
同前愛, 小松正明, 町野英徳, 中村康平, 金子修三, 生水貫人, 浅田健, 千代田達幸, 中山健太郎, 青木大輔, 京哲, 浜本隆二
Organizer
第79回日本癌学会学術総会
Related Report
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