NanoCluster Beacons Enable Detection of a Single N6-Methyladenine
NanoCluster Beacons Enable Detection of a Single N6-Methyladenine
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DOI：
10.1021/jacs.5b06038
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发表时间：
2015-08-26
影响因子：
15
通讯作者：
Yeh, Hsin-Chih
中科院分区：
文献类型：
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作者：
Chen, Yu-An;Obliosca, Judy M.;Yeh, Hsin-Chih
虽然N-6-甲基腺嘌呤（m（6）A）是原核生物和低等真核生物基因组中常见的修饰，并且具有许多生物学功能，但是没有简单且成本有效的方法来鉴定核酸靶标中的单个N-6-甲基腺嘌呤。本文介绍了一种基于银簇探针的甲基腺嘌呤特异性纳米簇信标（maNCB）的方法，该方法基于银簇的荧光发射光谱，可以检测DNA靶点中的单个m（6）A。maNCB不仅可以在单碱基水平上识别m（6）A，而且还可以定量异质样品中腺嘌呤甲基化的程度。我们的方法是上级的高分辨率熔融分析，因为我们可以精确定位的目标中的m（6）A的位置。
While N-6-methyladenine (m(6)A) is a common modification in prokaryotic and lower eukaryotic genomes and has many biological functions, there is no simple and cost-effective way to identify a single N-6-methyladenine in a nucleic acid target. Here we introduce a robust, simple, enzyme-free and hybridization-based method using a new silver cluster probe, termed methyladenine-specific NanoCluster Beacon (maNCB), which can detect single m(6)A in DNA targets based on the fluorescence emission spectra of silver clusters. Not only can maNCB identify m(6)A at the single-base level but it also can quantify the extent of adenine methylation in heterogeneous samples. Our method is superior to high-resolution melting analysis as we can pinpoint the location of m(6)A in the target.