Identification of methylated DNA-binding RNA aptamer and development of high sensitive DNA methylation detection system

甲基化DNA结合RNA适体的鉴定及高灵敏DNA甲基化检测系统的开发

• 批准号: 24760647
• 负责人: YOSHIDA Wataru
• 金额: $ 2.91万
• 依托单位: Tokyo University of Technology (2013)Tokyo University of Agriculture and Technology (2012)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-24760647/
• 关键词: アプタマー; in vitro selection; メチル化DNA; ＤＮＡ methylation; Ａｐｔａｍｅｒ

The research project aimed to identify RNA aptamer that specifically recognizes methylated DNA and develop high sensitive detection system for DNA methylation on target gene. To identify the aptamer, in vitro selection was performed against methylated duplex DNA. After 5th round selection, the RNA library was enriched. By sequencing analysis of the enriched RNA library, four kinds of sequence were identified in the library. The RNAs were synthesized and then binding assay against methylated DNA was performed. As the results, the RNAs bound to both of the methylated duplex DNA and unmethylated duplex DNA with 10 nM Kd. Therefore, it is expected that DNA methylation-binding RNA aptamer might be obtained by improving the binding specificity of the identified RNA aptamer and it might contribute for development of the high sensitive DNA methylation detection system.

本研究旨在寻找特异性识别甲基化DNA的RNA适配子，并建立高灵敏度的靶基因DNA甲基化检测体系。为了鉴定适配子，对甲基化的双链DNA进行了体外筛选。经第5轮筛选后，RNA文库得到丰富。通过对该文库的测序分析，确定了该文库中的4种序列。合成RNA，进行与甲基化DNA的结合实验。结果表明，甲基化的双链DNA和未甲基化的双链DNA均以10 nM kD结合。因此，通过提高已鉴定的RNA适配子的结合特异性，有望获得DNA甲基化结合的RNA适配子，为建立高灵敏度的DNA甲基化检测体系奠定基础。

项目成果

Aptamer selection based on genomic information; focusing on G-quadruplex forming sequence

基于基因组信息的适体选择；

• 发表时间: 2013
• 作者: Taiki Saito;Wataru Yoshida;Tomomi Yokoyama;Kazunori Ikebukuro
• 通讯作者: Kazunori Ikebukuro

Aptamer selection based on putative G-quadruplex sequence in genome

基于基因组中假定的 G-四链体序列的适体选择

• 发表时间: 2013
• 作者: Taiki Saito;Wataru Yoshida;Tomomi Yokoyama;Kentaro Teramoto;Kazunori Ikebukuro
• 通讯作者: Kazunori Ikebukuro

特異的蛍光リガンドとDNAマイクロアレイを基盤とした新規グアニン四重鎖形成配列の網羅的検出法の開発

基于特异性荧光配体和DNA微阵列的新型鸟嘌呤四链体形成序列综合检测方法的开发

• 发表时间: 2013
• 作者: 飯田圭介;寺正行;広川貴次;新家一男;長澤和夫
• 通讯作者: 長澤和夫

VEGF結合アプタマーのin silico maturation

VEGF 结合适体的计算机成熟

• 发表时间: 2012
• 作者: 安藤満;高橋有己;西川元也;渡部好彦;高倉喜信;野中 芳彦・深谷 剛弘・阿部 公一・ 吉田 亘・池袋 一典
• 通讯作者: 野中 芳彦・深谷 剛弘・阿部 公一・ 吉田 亘・池袋 一典

In silicoアプタマー探索法の開発

计算机适体搜索方法的开发

• 发表时间: 2012
• 作者: 吉田亘;齊藤大希;池袋一典
• 通讯作者: 池袋一典