Cellular microRNA detection with miRacles: microRNA-activated conditional looping of engineered switches
Cellular microRNA detection with miRacles: microRNA-activated conditional looping of engineered switches
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DOI：
10.1126/sciadv.aau9443
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发表时间：
2019-03-01
期刊：
影响因子：
13.6
通讯作者：
Halvorsen, Ken
中科院分区：
文献类型：
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作者：
Chandrasekaran, Arun Richard;MacIsaac, Molly;Halvorsen, Ken
MicroRNAs是一种短小的非编码调控RNA，越来越多地被用作疾病的生物标志物。检测微小RNA可能是困难和昂贵的，通常需要扩增、标记或放射性探针。在这里，我们报告了一种使用构象响应型DNA纳米开关的单步、非酶微RNA检测方法。我们的方法被称为奇迹(MicroRNA激活的工程开关条件性环路)，使用琼脂糖凝胶电泳法具有亚致敏的敏感性和单核苷酸特异性。我们从分化肌肉细胞的纳克级RNA提取液中检测到细胞microRNAs，并对一个生物样本中的几个microRNAs进行了多重检测。我们无需昂贵的设备或试剂即可实现1小时的检测，这使得该检测方法成为定量聚合酶链式反应和Northern杂交的有力替代。
MicroRNAs are short noncoding regulatory RNAs that are increasingly used as disease biomarkers. Detection of microRNAs can be arduous and expensive and often requires amplification, labeling, or radioactive probes. Here, we report a single-step, nonenzymatic microRNA detection assay using conformationally responsive DNA nanoswitches. Termed miRacles (microRNA-activated conditional looping of engineered switches), our assay has subattomole sensitivity and single-nucleotide specificity using an agarose gel electrophoresis readout. We detect cellular microRNAs from nanogram-scale RNA extracts of differentiating muscle cells and multiplex our detection for several microRNAs from one biological sample. We demonstrate 1-hour detection without expensive equipment or reagents, making this assay a compelling alternative to quantitative polymerase chain reaction and Northern blotting.