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Please use this identifier to cite or link to this item: http://hdl.handle.net/10119/14323

Title: 遺伝子修復治療を目指した化学的RNA Editing法の確立
Other Titles: Establishment of chemical RNA editing method for genetic code resoration therapy
Authors: 塚原, 俊文
Authors(alternative): Tsukahara, Toshifumi
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
Issue Date: 7-Jun-2017
Abstract: BFP-mRNAに対して脱アミノ化誘起用ODNを用いて光化学的な部位特異的脱アミノ化処理を行った。光化学的RNA editingは、RT-PCR-RFLPとin vitro翻訳したタンパク質の蛍光を確認した。また、光化学的RNA editingした当該mRNAの全長をシークエンスし、他の箇所に変異が生じていないことも確認した。効率的な脱アミノ化を誘起するODNデザインについても検討し、相補的配列長が14塩基で、ヘアピン長が9塩基のODNが12.4%と最も高い光化学的RNA editing効率を示した。この結果は光化学的RNA editing法が疾患治療法として適用可能であることを示唆している。: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.
Description: 基盤研究(B)(一般)
研究期間:2013~2016
課題番号:25290072
研究者番号:60207339
研究分野:分子生物学
Language: jpn
URI: http://hdl.handle.net/10119/14323
Appears in Collections:2016年度 (FY 2016)

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