Single-Molecule Sensor for High-Confidence Detection of miRNA.
Single-Molecule Sensor for High-Confidence Detection of miRNA.
复制标题
单分子传感器，用于高信心检测miRNA。
DOI：
10.1021/acssensors.1c02748
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发表时间：
2022-04-22
期刊：
影响因子：
8.9
通讯作者：
Dhakal, Soma
中科院分区：
文献类型：
--
作者：
Wijesinghe, Kalani M.;Kanak, Mazhar A.;Harrell, J. Chuck;Dhakal, Soma
关键词：
microRNAs（miRNAs）在调节基因表达中起着至关重要的作用，并与许多疾病有关。因此，灵敏和准确地检测疾病相关的miRNA对于疾病早期诊断的新兴革命至关重要。虽然由于其固有特性，如小尺寸、miRNA之间的高序列相似性和生物流体中的低丰度，miRNA的检测是一个挑战，但大多数miRNA检测策略涉及靶/信号扩增或涉及复杂的传感设计。在这项研究中，我们开发并测试了一种基于DNA的荧光共振能量转移（FRET）传感器，该传感器能够超灵敏地检测三阴性乳腺癌（TNBC）细胞表达的miRNA生物标志物（miRNA-342- 3 p）。该传感器显示了一个相对较低的FRET状态在没有一个目标，但连续的FRET转换之间的低和高FRET状态的存在下的目标被观察到。该传感器具有高度特异性，具有低至低飞摩尔（fM）的检测限，而无需放大目标，并且具有扩展至300，000 fM的大动态范围（3个数量级）。使用这种策略，我们证明了传感器允许检测来自癌细胞系和TNBC患者来源的异种移植物（PDX）的miRNA提取物中的miRNA-342- 3 p。由于设计简单，基于杂交的检测，这里开发的传感平台可用于检测各种各样的miRNA，从而能够早期诊断和筛选其他遗传疾病。
MicroRNAs (miRNAs) play a crucial role in regulating gene expression and have been linked to many diseases. Therefore, sensitive and accurate detection of disease-linked miRNAs is vital to the emerging revolution in early diagnosis of diseases. While the detection of miRNAs is a challenge due to their intrinsic properties such as small size, high sequence similarity among miRNAs and low abundance in biological fluids, the majority of miRNA-detection strategies involve either target/signal amplification or involve complex sensing designs. In this study, we have developed and tested a DNA-based fluorescence resonance energy transfer (FRET) sensor that enables ultrasensitive detection of a miRNA biomarker (miRNA-342-3p) expressed by triple-negative breast cancer (TNBC) cells. The sensor shows a relatively low FRET state in the absence of a target, yet continuous FRET transitions between low- and high-FRET states in the presence of the target are observed. The sensor is highly specific, has a detection limit down to low femtomolar (fM) without having to amplify the target, and has a large dynamic range (3 orders of magnitude) extending to 300,000 fM. Using this strategy, we demonstrated that the sensor allows detection of miRNA-342-3p in the miRNA-extracts from cancer cell lines and TNBC patient-derived xenografts (PDX). Given the simple-to-design hybridization-based detection, the sensing platform developed here can be used to detect a wide range of miRNAs enabling early diagnosis and screening of other genetic disorders.
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