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用于微RNA检测的i-Motif DNA分子信标

Tengshuo Luo^1,2, Ziyue Guan^1,2, Yunlong Liu¹

¹Department of Biomedical Engineering, School of Engineering, China Pharmaceutical University, Nanjing 211198, China.

Nucleic acids research

|June 26, 2025

概括

我们开发了一种新型的DNA探针,即i-motif分子信标 (iMB),用于敏感的核酸检测. 这种iMB通过DSN辅助的放大策略提供可控制的特异性和增强的灵敏度.

科学领域:

生物化学生物化学
分子生物学分子生物学
橄核酸的化学成分

背景情况:

分子信标 (MBs) 是用于核酸检测的寡核酸探针.
传统的MB依赖于光信号的茎环结构.
需要新的MB设计,具有增强的灵敏度和可控制的特性.

研究的目的:

为核酸检测引入一种新的基于i-motif DNA的分子信标 (iMB).
研究iMB的结合性质,并优化iMB的检测条件.
使用双重特定核酶 (DSN) 辅助放大策略来增强iMB的灵敏度.

主要方法:

基于i-motif DNA的分子信标 (iMB) 的设计和合成.
使用不同pH值和温度的相位图分析iMB的结合特性.
实施DSN辅助的放大策略,以提高灵敏度.
在细胞系和患者血中检测微RNA (miRNA),通过RT-qPCR验证.

主要成果:

在目标杂交时,iMB会选择性地打开,形成双重.
使用pH值和温度优化了结合特性和检测条件.
由于i-motif结构,iMB表现出可控制的灵敏度和特异性.
通过DSN辅助的放大,增加了超过四个数量级的灵敏度.
在生物样本中成功检测出miRNAs.

结论:

对于MBs来说,i-motif结构提供了对pH和温度敏感的茎.
开发的iMB为敏感和特定的核酸检测提供了一个多功能平台.
这种方法扩大了分子信标的结构多样性,用于各种应用.

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