Analysis of mechanisms of circular RNA translation in eukaryotic systems

• Project/Area Number: 18K14357
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 37030:Chemical biology-related
• Research Institution: Nagoya University
• Principal Investigator: Abe Naoko 名古屋大学, 理学研究科, 特任助教 (70772119)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 環状RNA / 翻訳反応 / RNA / 環状構造 / 真核細胞翻訳系 / 環状 / 翻訳

Outline of Final Research Achievements

1. Sequence dependence of the circular RNA translation was revealed. 2. It was shown that the translation efficiency of linear mRNA having a pseudocyclic structure was improved. 3. A synthetic method of chemically linking circular RNA was examined, and it was shown that mRNA containing unnatural linkage could be translated. 4. The mRNA in which the phosphorothioate modification was introduced in the 5'untranslated region in a limited manner greatly improved the translation activity in the E. coli cell-free translation system. This indicates that the introduction of position-specific chemical modification is an effective method for improving the translational activity of RNA.

Academic Significance and Societal Importance of the Research Achievements

従来起こりにくいと考えられてきた環状RNAの翻訳反応が起こりうること、塩基配列が翻訳効率に与える影響について検証し明らかにした。mRNAを医薬品として用いるに際しその安定性向上や翻訳能向上が必要であり、そのための方法論を多角的に検証した。その結果、今後の核酸医薬品の開発に役立つ知見を得た。

Research Products

• [Journal Article] Intracellular Delivery of Antisense DNA and siRNA with Amino Groups Masked with Disulfide Units (2020)
  • Author(s): Shu Zhaoma、Ota Azumi、Takayama Yukiya、Katsurada Yuri、Kusamori Kosuke、Abe Naoko、Nakamoto Kosuke、Tomoike Fumiaki、Tada Seiichi、Ito Yoshihiro、Nishikawa Makiya、Kimura Yasuaki、Abe Hiroshi
  • Journal Title: Chemical and Pharmaceutical Bulletin Volume: 68 Issue: 2 Pages: 129-132
  • DOI: 10.1248/cpb.c19-00811
  • NAID: 130007793432
  • ISSN: 0009-2363, 1347-5223
  • Year and Date: 2020-02-01
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Chemically synthesized circular RNAs with phosphoramidate linkages enable rolling circle translation (2020)
  • Author(s): Nakamoto Kosuke、Abe Naoko、Tsuji Genichiro、Kimura Yasuaki、Tomoike Fumiaki、Shimizu Yoshihiro、Abe Hiroshi
  • Journal Title: Chemical Communications Volume: - Issue: 46 Pages: 6217-6220
  • DOI: 10.1039/d0cc02140g
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Phosphorothioate Modification of mRNA Accelerates the Rate of Translation Initiation to Provide More Efficient Protein Synthesis (2020)
  • Author(s): Daisuke Kawaguchi, Ayumi Kodama, Naoko Abe, Kei Takebuchi, Fumitaka Hashiya, Fumiaki Tomoike, Kosuke Nakamoto, Yasuaki Kimura, Yoshihiro Shimizu, and Hiroshi Abe
  • Journal Title: Angewandte Chemie International Edition Volume: 59 Issue: 40 Pages: 17403-17407
  • DOI: 10.1002/anie.202007111
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Intracellular build-up RNAi with single-strand circular RNAs as siRNA precursors (2020)
  • Author(s): Kimura Yasuaki、Shu Zhaoma、Ito Mika、Abe Naoko、Nakamoto Kosuke、Tomoike Fumiaki、Shuto Satoshi、Ito Yoshihiro、Abe Hiroshi
  • Journal Title: Chemical Communications Volume: 56 Issue: 3 Pages: 466-469
  • DOI: 10.1039/c9cc04872c
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Translational control by secondary-structure formation in mRNA in a eukaryotic system (2020)
  • Author(s): Daisuke Kawaguchi, Saaya Shimizu, Naoko Abe, Fumitaka Hashiya, Fumiaki Tomoike,Yasuaki Kimura, Hiroshi Abe
  • Journal Title: Nucleosides, Nucleotides and Nucleic Acids Volume: 39 Issue: 1-3 Pages: 195-203
  • DOI: 10.1080/15257770.2019.1671593
  • Peer Reviewed
• [Journal Article] N6-methyl adenosine in siRNA evades immune response without reducing RNAi activity (2019)
  • Author(s): Akihiro Imaeda, Fumiaki Tomoike, Mayu Hayakawa, Kosuke Nakamoto, Yasuaki Kimura, Naoko Abe, Hiroshi Abe
  • Journal Title: Nucleosides, Nucleotides and Nucleic Acids Volume: 38 Issue: 12 Pages: 972-979
  • DOI: 10.1080/15257770.2019.1641205
  • Peer Reviewed
• [Journal Article] Disulfide-Unit Conjugation Enables Ultrafast Cytosolic Internalization of Antisense DNA and siRNA (2019)
  • Author(s): Shu Zhaoma、Tanaka Iku、Ota Azumi、Fushihara Daichi、Abe Naoko、Kawaguchi Saki、Nakamoto Kosuke、Tomoike Fumiaki、Tada Seiichi、Ito Yoshihiro、Kimura Yasuaki、Abe Hiroshi
  • Journal Title: Angewandte Chemie International Edition Volume: 印刷中 Issue: 20 Pages: 6611-6615
  • DOI: 10.1002/anie.201900993
  • NAID: 130007841763
  • Peer Reviewed
• [Presentation] RNAのホスホロチオエート修飾による翻訳反応の促進 (2021)
  • Author(s): 阿部奈保子、川口大輔、児玉亜有実、橋谷文貴、木村康明、清水義宏、阿部洋
  • Organizer: 日本薬学会第１４１年会
• [Presentation] 翻訳反応サイクルを加速する人工mRNAの開発 (2020)
  • Author(s): 川口大輔、児玉亜有実、阿部奈保子、竹渕慧、橋谷文貴、友池史明、中本航介、木村康明、清水義宏、阿部洋
  • Organizer: 第１４回バイオ関連シンポジウム
• [Presentation] 化学修飾核酸の合成と核酸医薬への応用 (2020)
  • Author(s): 川口大輔、児玉亜有実、阿部奈保子、竹渕慧、橋谷文貴、友池史明、中本航介、木村康明、清水義宏、阿部洋
  • Organizer: 第10回CSJ化学フェスタ2020
• [Presentation] 二次構造形成によるmRNAの翻訳反応制御 (2019)
  • Author(s): 川口 大輔、清水 沙彩、阿部 奈保子、友池 史明、木村 康明、阿部 洋
  • Organizer: 日本ケミカルバイオロジー学会 第14回年会
• [Presentation] 二次構造形成によるmRNAの翻訳反応制御 (2019)
  • Author(s): 川口 大輔、清水 沙彩、阿部 奈保子、友池 史明、木村 康明、阿部 洋
  • Organizer: 日本核酸医薬学会第5回年会
• [Presentation] 真核生物における環状RNAを用いた終わりのない回転式翻訳現象 (2018)
  • Author(s): 清水 沙彩、児玉 亜有実、阿部 奈保子、友池 史明、木村 康明、阿部 洋
  • Organizer: 第20回日本RNA学会年会
• [Presentation] 真核生物における環状RNAを用いた終わりのない回転式翻訳現象 (2018)
  • Author(s): 阿部 奈保子、清水 沙彩、児玉 亜有実、友池 史明、木村 康明、阿部 洋
  • Organizer: 第12回バイオ関連化学シンポジウム
• [Presentation] Circular RNA for protein translation (2018)
  • Author(s): Naoko Abe, Hiroshi Abe
  • Organizer: FNA Perth 2018
  • Int'l Joint Research
• [Book] 核酸科学ハンドブック、第III部、6. 環状核酸 (2020)
  • Author(s): 阿部奈保子、阿部洋
  • Publisher: 講談社
  • ISBN: 9784065207864
• [Book] 化学 第７５巻第１０号 解説 翻訳反応サイクルを加速するmRNA分子 (2020)
  • Author(s): 川口大輔、阿部奈保子、阿部洋
  • Total Pages: 7
  • Publisher: 化学同人
• [Patent(Industrial Property Rights)] 切断プライマー (2020)
  • Inventor(s): 阿部洋、阿部奈保子、中本航介、村瀬裕貴、木村康明
  • Industrial Property Rights Holder: 阿部洋、阿部奈保子、中本航介、村瀬裕貴、木村康明
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2019-140851
  • Filing Date: 2020
• [Patent(Industrial Property Rights)] 停止プライマー (2020)
  • Inventor(s): 阿部洋、阿部奈保子、中本航介、村瀬裕貴、木村康明
  • Industrial Property Rights Holder: 阿部洋、阿部奈保子、中本航介、村瀬裕貴、木村康明
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2019-140852
  • Filing Date: 2020
• [Patent(Industrial Property Rights)] キャップ化ＲＮＡの製造方法 (2020)
  • Inventor(s): 阿部洋、木村康明、阿部奈保子
  • Industrial Property Rights Holder: 阿部洋、木村康明、阿部奈保子
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2020-032889
  • Filing Date: 2020