核糖核酸
劈理(地质)
核糖核酸酶P
化学
环介导等温扩增
核糖核酸酶H
计算生物学
分子生物学
组合化学
生物化学
生物
DNA
基因
断裂(地质)
古生物学
作者
Yuanyuan Sun,Fangfang Sun,Bing-Jie Han,Chenghui Liu
标识
DOI:10.1016/j.snb.2021.131200
摘要
Specific and accurate determination of N1-Methyladenosine (m1A) is of critical significance to better understand its biological functions. Precise quantification of low level of RNA modification against a high background of homologous normal RNA still remains a big challenge. To solve this issue, a facile site-specific cleavage-assisted isothermal exponential amplification reaction (SSC-EXPAR) is skillfully designed for direct m1A assay. By exploiting the highly selective RNA cleavage, the normal RNA can be disconnected to avoid the false-positive interference, while the intact m1A can mediate and trigger three-way junction structure-based EXPAR (3WJ-EXPAR). The elegant design of m1A-aroused positive signal can guarantee the accurate and reliable detection of low-level m1A. As low as 1 fM of m1A-containing RNA can be accurately determined with a specificity allowing the discrimination of 0.5% m1A against unmodified adenosine (A) at single-base resolution. In virtue of the outstanding sensitivity and specificity, the SSC-EXPAR approach holds great potential in further understanding of m1A-associated biofunctions and the clinical value in diagnosis and therapeutic treatment of human cancers.
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