Light-up Hoechst-DNA aptamer pair: Generation of an aptamer-selective fluorophore from a conventional DNA-staining dye
Light-up Hoechst-DNA aptamer pair: Generation of an aptamer-selective fluorophore from a conventional DNA-staining dye
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DOI：
10.1002/cbic.200700325
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发表时间：
2007-10-15
期刊：
影响因子：
3.2
通讯作者：
Aoyama, Yasuhiro
中科院分区：
文献类型：
--
作者：
Sando, Shinsuke;Narita, Atsushi;Aoyama, Yasuhiro
我们已经设计了一种策略，以基于常规DNA染色染料的向下修饰来产生发光荧光团-适体对，以抑制其对原始dsDNA靶标的亲和力，然后重新选择将结合至经修饰的染料的适体。根据这条线，我们制备了一种微极性敏感的Hoechst衍生物，具有两个tBu基团，对通常富含AT的dsDNA靶标具有低亲和力。在体外从随机池中选择的DNA适体作为触发器，以增强否则非荧光Hoechst衍生物的荧光，缩短的25聚体序列显示出显着的增强（点亮）。将25-mer序列分成二元适体探针，从而使我们能够通过使用未修饰的DNA作为探针以单核苷酸分辨率检测靶核酸序列。
We have designed a strategy to generate a light-up fluorophore-aptamer pair based on a down-modification of a conventional DNA-staining dye to suppress its affinity to the original dsDNA targets, followed by reselection of aptamers that would bind to I the modified dye. Following this line, we prepared a micropolarity-sensitive Hoechst derivative possessing two tBu groups with low affinity to the usual AT-rich dsDNA targets. DNA aptamers selected in vitro from a random pool worked as triggers to enhance the fluorescence of an otherwise nonfluorescent Hoechst derivative, and the shortened 25-mer sequence showed remarkable enhancement (light-up). The 25-mer sequence was split into binary aptamer probes, thus enabling us to detect a target nucleic acid sequence with a single-nucleotide resolution by use of unmodified DNA as a probe.