Establishment of chemical RNA editing method for genetic code resoration therapy

2016 Fiscal Year Final Research Report

• Project/Area Number: 25290072
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Medical genome science
• Research Institution: Japan Advanced Institute of Science and Technology
• Principal Investigator: Tsukahara Toshifumi 北陸先端科学技術大学院大学, 先端科学技術研究科, 教授 (60207339)
• Co-Investigator(Kenkyū-buntansha): 鈴木 仁 北陸先端科学技術大学院大学, マテリアルサイエンス研究科, 助教 (00447690) ; 藤本 健造 北陸先端科学技術大学院大学, マテリアルサイエンス研究科, 教授 (90293894)
• Co-Investigator(Renkei-kenkyūsha): Takada Shin'ichi 国立研究開発法人国立精神・神経医療研究センター, 神経研究所, 所長 (90171644)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Keywords: 疾患治療 / 点変異 / RNA editing / 脱アミノ化 / 蛍光タンパク質 / 遺伝コード修復 / RFLP / 恒常的発現細胞

Outline of Final Research Achievements

Photochemical site-specific deamination was performed toward BFP-mRNA, by using oligodeoxynucleotides,ODNs, for inducing deamination. The base substitution by artificial RNA editing was confirmed by RT-PCR-RFLP analysis and fluorescence of in-vitro-translated proteins. Moreover, the mRNA sample was sequenced after photochemical RNA editing treatments, and confirmed that there was no mutation occurred except the target-site. We also investigated the ODN design that could induce efficient deamination. The ODN with complementary sequence length of 14 bases and hairpin length of 9 bases showed the highest photochemical RNA editing efficiency at 12.4%. This result suggests that the photochemical RNA editing is applicable as a theraputic method for genetic diseases.

Free Research Field

分子生物学