{"title":"Simultaneous sensitive detection and imaging of dual microRNAs through DNA tetrahedral scaffold-corbelled autonomous-motion molecular machine.","authors":"Yun Zhang, Liang Gao, Zhe Shi, Qiong Wu, Xiangmin Miao","doi":"10.1016/j.talanta.2025.127556","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"286 ","pages":"127556"},"PeriodicalIF":5.6000,"publicationDate":"2025-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Talanta","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1016/j.talanta.2025.127556","RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
引用次数: 0
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.
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.