Development of modified U1 snRNA derivatives as new nucleic acid drugs

• Project/Area Number: 16K05840
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bio-related chemistry
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: Ohkubo Akihiro 東京工業大学, 生命理工学院, 准教授 (60376960)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: U snRNA / スプライシング / 核酸合成 / U1 snRNA / U11 snRNA / 核酸医薬 / 核酸

Outline of Final Research Achievements

In this study, we carried out synthesis of a U snRNA by the ligation reactions using four RNA oligomers and RNA ligase. However, the ligation efficiency was very low due to the steric hindrance of the RNA structure. In order to overcome the steric hindrance, we used DNA ligase in the presence of a splint DNA. As a result, we successfully synthesize a U snRNA in an efficient yield.
Moreover, we also synthesized modified U1 and U11 snRNA analogs containing a m3G cap structure under the ligation conditions using DNA ligase in the presence of a splint DNA. At the present, the splicing activities of the U RNAs are evaluated cystic fibrosis model cells.

Academic Significance and Societal Importance of the Research Achievements

遺伝子変異によるエキソンの欠落を化学合成したRNAによって阻止する「スプライシングコレクション」は、これまで治療が困難とされてきたスプライシング異常に起因する遺伝子疾患の治療に多いに貢献できると期待される。また、従来の核酸医薬とは全く違った作用機序をもつため、当該分野における貢献度も高いと考えられる。

Research Products

• [Journal Article] Synthesis of and triplex formation in oligonucleotides containing 2’-deoxy-6-thioxanthosine (2018)
  • Author(s): akeshi Inde, Shuhei Nishizawa, Yuusaku Hattori, Takashi Kanamori, Hideya Yuasa, Kohji Seio, Mitsuo Sekine, Akihiro Ohkubo
  • Journal Title: Bioorg. Med. Chem. Volume: 26 Issue: 13 Pages: 3785-3790
  • DOI: 10.1016/j.bmc.2018.06.004
  • Peer Reviewed
• [Journal Article] pH-Dependent Switching of Base Pairs Using Artificial Nucleobases with Carboxyl Groups (2018)
  • Author(s): Kaewsomboon T, Nishizawa S, Kanamori T, Yuasa H, Ohkubo A
  • Journal Title: The Journal of Organic CHemistry Volume: 83 Issue: 3 Pages: 1320-1320
  • DOI: 10.1021/acs.joc.7b02828
  • Peer Reviewed
• [Journal Article] Fluorescence enhancement of oligodeoxynucleotides modified with green fluorescent protein chromophore mimics upon triplex formation (2017)
  • Author(s): Kanamori T, Takamura A, Tago N, Masaki Y, Ohkubo A, Sekine M, Seio K.
  • Journal Title: Organic and Biomolecular Chemistry Volume: 15 Issue: 5 Pages: 1190-1197
  • DOI: 10.1039/c6ob01278g
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Coating lanthanide nanoparticles with carbohydrate ligands elicits affinity for HeLa and RAW264.7 cells, enhancing their photodamaging effect (2017)
  • Author(s): Takashi Kanamori, Takashi Sawamura, Tatsumi Tanaka, Izumi Sotokawa, Ryota Mori, Kotaro Inada, Akihiro Ohkubo, Shun-Ichiro Ogura, Yasutoshi Murayama, Eigo Otsuji, Hideya Yuasa
  • Journal Title: Bioorganic & Medicinal Chemistry Volume: 25 Issue: 2 Pages: 743-749
  • DOI: 10.1016/j.bmc.2016.11.050
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Enzymatic synthesis and reverse transcription of RNAs incorporating 2 -O-carbamoyl uridine triphosphate (2016)
  • Author(s): Masaki Y, Ito H, Oda Y, Yamazaki K, Tago N, Ohno K, Ishii N, Tsunoda H, Kanamori T, Ohkubo A, Sekine M, Seio K.
  • Journal Title: Chemical Communications Volume: 52 Issue: 87 Pages: 12889-12892
  • DOI: 10.1039/c6cc05796a
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Synthesis and properties of triplex-forming oligonucleotides containing modified sugar moieties and nucleobases. (2018)
  • Author(s): S. Nishizawa, T. Ohnishi, L. Watanabe, T. Kanamori, H. Yuasa, A. Ohkubo
  • Organizer: The 45th International Symposium on Nucleic Acids Chemistry
  • Int'l Joint Research
• [Presentation] Synthesis and properties of cyclic oligonucleotides containing acyl groups at the 5'- and 3'- terminal sites. (2018)
  • Author(s): R. Hashimoto, S. Nishizawa, Y. Miyake, T. Kanamori, H. Yuasa, A. Ohkubo
  • Organizer: The 45th International Symposium on Nucleic Acids Chemistry
  • Int'l Joint Research
• [Presentation] U1 snRNA 誘導体の合成 (2017)
  • Author(s): 細谷輝、金森功吏、湯浅 英哉、大窪章寛
  • Organizer: 日本薬学会春季年会
  • Place of Presentation: 仙台国際センター
  • Year and Date: 2017-03-25
• [Presentation] イソフタル酸誘導体を有する新規修飾核酸の合成と 性質 (2017)
  • Author(s): 西村ゆり、Kaewsomboon TANASAK、金森功吏、湯浅 英哉、大窪章寛
  • Organizer: 日本薬学会春季年会
  • Place of Presentation: 仙台国際センター
  • Year and Date: 2017-03-25
• [Presentation] 5´および3´にアシル基を有する環状オリゴヌクレオチドの合成 (2017)
  • Author(s): 西澤周平、橋本律、金森 功吏、湯浅英哉、大窪章寛
  • Organizer: 日本化学会 春季年会
  • Place of Presentation: 慶応大学
  • Year and Date: 2017-03-19
• [Presentation] Synthesis and properties of cyclic oligonucleotides containing acyl groups at the 5´- and 3´-terminal sites. (2017)
  • Author(s): Tanasak kaewsomboon, Shuhei Nishizawa, Ritsu Hashimoto, Yu Miyake, Takashi Kanamori, Hideya Yuasa, Akihiro Ohkubo
  • Organizer: International Symposium of Nucleic Acids Chemistry
  • Int'l Joint Research
• [Presentation] Synthesis and properties of new modified oligonucleotides containing amino groups at the 5´-positions of ribose moieties (2017)
  • Author(s): Tomohiro Aoyama, Kosuke Muto, Lintaro Watanabe, Takashi Kanamori, Hideya Yuasa, Akihiro Ohkubo
  • Organizer: International Symposium of Nucleic Acids Chemistry
  • Int'l Joint Research