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基于碳点和分子信标的荧光传感器用于检测癌症标志物 microRNA-21。

A carbon dot and molecular beacon based fluorometric sensor for the cancer marker microRNA-21.

机构信息

Department of Chemistry, Faculty of Chemistry and Chemical Engineering, Graduate University of Advanced Technology, Kerman, 7631818356, Iran.

Department of Nanotechnology, Faculty of Sciences and Modern Technologies, Graduate University of Advanced Technology, Kerman, 7631818356, Iran.

出版信息

Mikrochim Acta. 2019 Feb 1;186(3):132. doi: 10.1007/s00604-019-3233-z.

Abstract

A carbon quantum dot (CQD) labeled molecular beacon was synthesized and applied to the detection of microRNA-21. The CQDs possess low cytotoxicity, excellent water solubility, and photostability. The CQDs were characterized by transmission electron microscopy, dynamic light scattering, Fourier-transform infrared spectroscopy, and fluorescence spectroscopy. The molecular beacon (MB) was labeled with the CQDs at the 5' end, and with Black Hole Quencher 1 (BHQ1) at the 3' end. The two labels act as the donor and acceptor parts of a FRET system, respectively. Only weak fluorescence is observed in the absence of microRNA-21, and in the presence of scrambled or mismatched sequences. However, in the presence of microRNA-21, fluorescence intensity of the CQDs at 460 nm (excitation at 360 nm) recovers. The hybridization of the hairpin structure of the MB with microRNA-21 opens the loop of MB. Consequently, the distance between the BHQ1 quencher and the CQDs is increased and fluorescence changes. The probe has high sensitivity (with a 0.3 nM limit of detection) and specificity. It can distinguish between microRNA-21 and its single mismatch mutant and hence represents a valuable tool for the early cancer diagnosis. Graphical abstract Schematic presentation of a fluorometric microR-21 assay using carbon dots carrying a molecular beacon (MB) labeled with a black hole quencher. Quenching is suppressed once the MB binds to microRNA-21.

摘要

一种碳量子点(CQD)标记的分子信标被合成并应用于 microRNA-21 的检测。CQD 具有低细胞毒性、优异的水溶性和光稳定性。通过透射电子显微镜、动态光散射、傅里叶变换红外光谱和荧光光谱对 CQD 进行了表征。分子信标(MB)在 5' 端用 CQD 标记,在 3' 端用黑洞猝灭剂 1(BHQ1)标记。这两个标签分别作为荧光共振能量转移(FRET)系统的供体和受体部分。在不存在 microRNA-21 的情况下,或者在存在乱序或错配序列的情况下,只能观察到微弱的荧光。然而,在存在 microRNA-21 的情况下,CQD 在 460nm 处的荧光强度(在 360nm 处激发)恢复。MB 的发夹结构与 microRNA-21 的杂交打开了 MB 的环。因此,BHQ1 猝灭剂和 CQD 之间的距离增加,荧光发生变化。该探针具有高灵敏度(检测限为 0.3nM)和特异性。它可以区分 microRNA-21 和其单碱基错配突变体,因此代表了癌症早期诊断的有价值的工具。

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