Development of the new method for estimate of radiation effect using a fluorescent modified oligonucleotide

• Project/Area Number: 16K18347
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Nuclear engineering
• Research Institution: University of Fukui
• Principal Investigator: MATUO Youichirou 福井大学, 学術研究院工学系部門, 講師 (90568883)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2018: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2017: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2016: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: 放射線 / DNA切断 / 蛍光修飾 / オリゴヌクレオチド / 生体影響 / 放射線影響 / 放射線防護 / 線量評価 / 放射線生物学 / 保健物理 / 原子力安全

Outline of Final Research Achievements

We are developing a direct measurement method to verify radiation damage on the bio-material employing an fluorescence modified oligonucleotide. The fluorescence modified oligonucleotide has a fluorescence site (6-FAM) and quenching site (TAMRA) on both terminals.We synthesized fluorescence modified 27 mer oligonucleotide of yeast URA3 and poly Adenine sequence. The samples were irradiated with helium ion beam at the Wakasawan Energy Research Center. The samples were also irradiated with 60Co gamma-rays at Osaka Univ. for the comparison with helium ion beam irradiated samples. In a present study we found that the fluorescence intensities increased with increase of absorbed dose of the helium ion beam and gamma-ray. Fluorescence intensity has been increased with a function of yield of molecular damage. Moreover, it has been suggested that the sensitivity may change depending on the sequence.

Academic Significance and Societal Importance of the Research Achievements

放射線による生体分子の損傷量を、損傷に起因する蛍光にて評価するものである。このことから、従来法にない以下のような特徴を有する。
①検出感度について、電気泳動後のゲル中の蛍光を測定する手法と比較して、直接分子鎖の切断を蛍光を介して計測する本手法は優れると考えられる。②前処理を評価とせず、蛍光強度の測定に必要な時間も数分程度であり、処理時間が短い。③配列を自由に設計できる。学術的効果の延長上として、放射線による生体影響研究のツールとして活用できる可能性がある。

Research Products

• [Journal Article] Biological effects of carbon ion beams with various LETs on budding yeast Saccharomyces cerevisiae (2018)
  • Author(s): Matuo Youichirou、Izumi Yoshinobu、Furusawa Yoshiya、Shimizu Kikuo
  • Journal Title: Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis Volume: 810 Pages: 45-51
  • DOI: 10.1016/j.mrfmmm.2017.10.003
  • Peer Reviewed
• [Journal Article] 放射線照射によるDNA損傷の新評価手法の検討 (2018)
  • Author(s): 松尾 陽一郎、平山 誠、川井 良太、砂川 武義、清水 喜久雄、泉 佳伸
  • Journal Title: 放射線生物研究 Volume: 53 Pages: 223-240
  • NAID: 120007146165
  • Peer Reviewed
• [Presentation] Development of the new method for estimate of radiation effect using a fluorescent-modifiedーoligonucleotide (2018)
  • Author(s): Ryota KAWAI, Youichirou MATUO, Takao KOJIMA, Kikuo SHIMIZU, Yoshinobu IZUMI
  • Organizer: 5th Asian and Oceanic Congress on Radiation Protection
  • Int'l Joint Research
• [Presentation] 蛍光修飾オリゴヌクレオチドを用いた放射線損傷評価手法の開発 (2018)
  • Author(s): 川井 良太，松尾 陽一郎，泉 佳伸
  • Organizer: 第61回放射線化学討論会, 日本放射線化学会若手の会
• [Presentation] 蛍光修飾ヌクレオチドを用いた放射線による生体分子の損傷量評価手法に関する基礎的研究 (2017)
  • Author(s): 松尾陽一郎，川井良太，泉佳伸，安田仲宏
  • Organizer: 第31回固体飛跡検出器研究会
  • Place of Presentation: 京都、日本
  • Year and Date: 2017-03-09
• [Presentation] 蛍光修飾を用いたオリゴヌクレオチドの放射線損傷評価に関する基礎的検討 (2017)
  • Author(s): 川井 良太、松尾 陽一郎、小嶋 崇夫、泉 佳伸
  • Organizer: 日本放射線安全管理学会 第16回学術大会
• [Presentation] Feasibility study on radiation-induced damage of DNAs using a fluorescence modified oligonucleotide (2017)
  • Author(s): Youichirou MATUO, Ryota KAWAI, Makoto, HIRAYAMA, Yoshinobu IZUMI, Nakahiro YASUDA
  • Organizer: 27 International Conference on Nuclear Tracks and Radiation Measurements
  • Int'l Joint Research