Copper-Free Click Chemistry-Mediated Assembly of Single Quantum Dot Nanosensor for Accurately Monitoring Locus-Specific m6A in Cancer Cells

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外文摘要：N6-methyladenosine(m6A)plays an important role in embryogenesis,nuclear export,transcription splicing,and protein translation control.Herein,we demonstrate a copper-free click chemistry-mediated assembly of single quantum dot(QD)nanosensor for accu-rately monitoring locus-specific m6A in cancer cells.The m6A-sensitive endoribonuclease MazF can digest the unmethylated A-RNA,and the intact m6A-RNA then hybridizes with DNA probes a and b to produce a sandwich hybrid,initiating the click chemistry to gen-erate probe a-b ligation product via first tandem ligation detection reaction(LDR)cycle.Subsequently,DNA probes c and d can hy-bridize with the probe a-b ligation product to generate the probe c-d ligation product via second LDR cycle.Both LDR cycles can be repeated through denaturation and annealing reaction to generate abundant biotin-/fluorophore-modified probe c-d ligation prod-ucts that can easily assemble on the QD surface to induce distinct fluorescence resonance energy transfer(FRET)between QD and Cy5.This assay can be homogenously performed without the involvement of copper catalyst,m6A-specific antibody,radioactive la-beling,ligase enzyme,enzymatic reverse transcription,and next-generation sequencing.Moreover,it can discriminate even 0.01％m6A level in complex samples and accurately measure cellular mA-RNA expression,providing a promising avenue for clinical diag-nostics and biomedical research.

外文关键词：

RNA methylationClick chemistryFörster resonance energy transferBiosensorCancer

作者：

Yue-Ying Li、Ning-Ning Zhao、Yi-Xuan Geng、Qian Han、Jian-Ge Qiu、Bing-Hua Jiang、Zi-Yue Wang、Chun-Yang Zhang

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作者单位：

Translational Medicine Center,The First Affiliated Hospital of Zhengzhou University,The Academy of Medical Sciences,Henan Institute of Medical and Pharmaceutical Sciences BGI College,Zhengzhou University,Zhengzhou,Henan 450052,China

College of Chemistry,Chemical Engineering and Materials Science,Shandong Normal University,Jinan,Shandong 250014,China

School of Chemistry and Chemical Engineering,Southeast University,Nanjing,Jiangsu 211189,China

基金：

国家自然科学基金

项目编号：

21735003

出版年：

2024

DOI：

10.1002/cjoc.202300595

中国化学(英文版)

中国化学会上海有机化学研究所

中国化学(英文版)

CSTPCD

影响因子：0.848

ISSN：1001-604X

年,卷(期)：2024.42(5)

参考文献量46