Simultaneous sensitive detection and imaging of dual microRNAs through DNA tetrahedral scaffold-corbelled autonomous-motion molecular machine.

IF 5.6 1区化学 Q1 CHEMISTRY, ANALYTICAL Talanta Pub Date : 2025-05-01 Epub Date: 2025-01-13 DOI:10.1016/j.talanta.2025.127556

Yun Zhang, Liang Gao, Zhe Shi, Qiong Wu, Xiangmin Miao

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Abstract

Sensitive and accurate detection and imaging of different microRNAs (miRNAs) in cancer cells hold great promise for early disease diagnosis. Herein, a DNA tetrahedral scaffold (DTS)-corbelled autonomous-motion (AM) molecular machine based fluorescent sensing platform was designed for simultaneous detection of two types of miRNAs (miRNA-21 and miRNA-155) in HeLa cells. Locking-strand-silenced DNAzymes (P:L duplex) were firstly grafted at the loop of target-analogue-embedded double-stem hairpin substrates (TDHS) of DTS, making the sensor in a "signal off" state due to the closely distance between modified fluorophores (FAM and Cy5) with the corresponding quenchers (BHQ1 and BHQ2). The detection of miRNA-21 and miRNA-155 was mainly based on the activation of locking-strand-silenced DNAzymes, cleaving hairpin DNA into single-strand DNA segments, accompanying with the release of modified fluorophores and the signal recovery (signal on). Upon the cyclical stimulation of miRNA targets in such AM molecular machine, sensitive detection of miRNA-21 and miRNA-155 was realized in this self-feedback circuit (SFC) with the detection limit down to 38.8 aM and 27.1 aM, respectively. Moreover, the analytical performance was greatly improved for miRNAs imaging in cancer cells with enhanced tumor cell recognition ability, excellent stability in virtue of DTS, indicating a potential analytical tool in early cancer diseases diagnosis.

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利用DNA四面体支架-骨架自主运动分子机同时灵敏检测和成像双microrna。

灵敏、准确地检测和成像癌细胞中不同的microrna （miRNAs）对早期疾病诊断具有很大的希望。本文设计了一种基于DNA四面体支架(DTS)-骨架自主运动（AM）分子机的荧光传感平台，用于同时检测HeLa细胞中的两种mirna （miRNA-21和miRNA-155）。锁定链沉默DNAzymes （P:L双工）首先接枝在DTS的目标模拟嵌入双茎发夹衬底（TDHS）的环路上，由于修饰的荧光团（FAM和Cy5）与相应的猝灭剂（BHQ1和BHQ2）之间的距离很近，使传感器处于“信号关闭”状态。miRNA-21和miRNA-155的检测主要是通过激活锁定链沉默的DNAzymes，将发夹DNA切割成单链DNA片段，同时释放修饰的荧光团，信号恢复（信号开启）。在AM分子机器中循环刺激miRNA靶点后，在自反馈电路（SFC）中实现了对miRNA-21和miRNA-155的灵敏检测，检测限分别降至38.8 AM和27.1 AM。同时，对肿瘤细胞miRNAs成像的分析性能也大大提高，具有增强的肿瘤细胞识别能力和良好的DTS稳定性，是一种潜在的癌症早期疾病诊断的分析工具。

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来源期刊

Talanta 化学-分析化学

CiteScore

12.30

自引率

4.90%

发文量

861

审稿时长

29 days

期刊介绍： Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome. Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.