A Ratiometric Fluorescent Bioprobe Based on Carbon Dots and Acridone Derivate for Signal Amplification Detection Exosomal microRNA.
A Ratiometric Fluorescent Bioprobe Based on Carbon Dots and Acridone Derivate for Signal Amplification Detection Exosomal microRNA.
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DOI：
10.1021/acs.analchem.8b01143
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发表时间：
2018-07
影响因子：
7.4
通讯作者：
Yaokun Xia;Liangliang Wang;Juan Li;Xiangqi Chen;Jianming Lan;An Yan;Yun Lei;Sheng Yang;
中科院分区：
文献类型：
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作者：
Yaokun Xia;Liangliang Wang;Juan Li;Xiangqi Chen;Jianming Lan;An Yan;Yun Lei;Sheng Yang;
最近，外泌体 microRNA (miRNA) 的灵敏和选择性检测受到了广泛关注，因为它与包括癌症在内的许多复杂疾病有关。在此，我们报道了一种基于 DNA 标记碳点 (DNA-CD) 和 5,7-二硝基-2-磺基吖啶酮 (DSA) 的比率荧光生物探针，与靶催化信号放大偶联，用于检测外泌体 miRNA-21。当生物探针组装时，碳点 (CD) 和 DSA 之间存在高荧光共振能量转移 (FRET) 效率。然而，在目标存在的情况下，随着荧光生物探针的拆卸，CD和DSA的荧光强度同时发生变化。由于双荧光强度的比率，这种比例荧光生物探针能够通过计算两个不同波长的发射强度比来抵消环境波动，对于外泌体 miRNA-21 的检测来说足够稳健和稳定。此外，我们还发现，理论上，单个 miRNA-21 可以通过 DSA 催化多个 CD 的分解，从而使检测 miRNA-21 的荧光比率产生显着变化。采用这种信号放大策略，检测限低至 3.0 fM。此外，由于引入锁核酸来介导链置换反应，该策略的选择性显着提高，甚至针对单碱基错配序列。更重要的是，我们的策略可以监测外泌体miRNA-21的动态变化，这可能成为区分癌症外泌体和非致瘤性外泌体的潜在工具。总之，这种比例荧光生物探针具有高稳定性、灵敏度和选择性耦合，操作简便且成本效益高，具有广泛应用的巨大潜力。
Recently, sensitive and selective detection of exosomal microRNAs (miRNAs) has been garnering significant attention, because it is related to many complex diseases, including cancer. Herein, we report a ratiometric fluorescent bioprobe based on DNA-labeled carbon dots (DNA-CDs) and 5,7-dinitro-2-sulfo-acridone (DSA) coupling with the target-catalyzing signal amplification for the detection of exosomal miRNA-21. There was high fluorescence resonance energy transfer (FRET) efficiency between carbon dots (CDs) and DSA when the bioprobe was assembled. However, in the presence of the target, with disassembling of the fluorescent bioprobe, the fluorescence intensities of CDs and DSA were changed simultaneously. Because of the ratio of dual fluorescence intensities, this ratiometric fluorescent bioprobe was able to cancel out environmental fluctuations by calculating emission intensity ratio at two different wavelengths, being robust and stable enough for detection of exosomal miRNA-21. In addition, we displayed that a single miRNA-21 can catalyze the disassembly of multiple CDs with DSA theoretically, yielding significant change in the fluorescence ratio for the detection of miRNA-21. With this signal amplification strategy, the limit of detection was as low as 3.0 fM. Furthermore, because of the introduction of lock nucleic acid to mediate the strand displacement reaction, the selectivity of this strategy was improved remarkably, even against single base mismatch sequence. More importantly, our strategy could monitor the dynamic change of exosomal miRNA-21, which maybe becomes a potential tool to distinguish cancer exosomes and nontumorigenic exosomes. In a short, this ratiometric fluorescence bioprobe possessed high stability, sensitivity and selectivity coupling with ease of operation and cost efficiency, leading to great potential for wide application.