A novel photoelectrochemical strategy for ultrasensitive and simultaneous detection of 5-methylcytosine and N6-methyladenosine based on proximity binding-triggered assembly MNAzyme -mediated HRCA

IF 5.3 2区化学 Q1 CHEMISTRY, ANALYTICAL Microchimica Acta Pub Date : 2025-02-26 DOI:10.1007/s00604-025-07033-z

Yue Hu, Mengshi Xia, Mimi Li, Lulu Li, Chenghong Li, Yi Liu, Lina Wang, Hui Huang, Lichao Fang, Kexing Peng, Huamin Liu, Xiaolong Wang, Junsong Zheng

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Abstract

The preparation of UiO-66@CdTe@AuNPs composites is presented for the first time, which function as a photoelectrochemical (PEC) sensing matrix and are conjugated to a three-way junction (TWJ). We propose an antibody-based specific recognition-induced neighbor-joining reaction that initiates the assembly of two molecularly designed nucleic acid enzymes (MNAzymes), to release an oligonucleotide that hydrolyzes TWJ through the mechanism of a toehold-mediated strand displacement reaction (TSDR). Subsequently, a hybridization chain reaction application (HRCA)-based dendrimer is formed, which immobilizes a large number of quantum dots to generate a burst effect that reduces the photocurrent signal. As anticipated, the developed PEC biosensor showed excellent analytical performance for both m⁶A-RNA and m⁵C-RNA, with detection limits of 0.309 fM and 6.918 aM, respectively. The successful fabrication of this ultrasensitive and simultaneous PEC biosensor provides new insights for epigenetic research and the bioassay, mechanism investigation and clinical diagnosis of diseases associated with RNA methylation.

Graphical Abstract

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基于近距离结合触发组装MNAzyme介导的HRCA的超灵敏和同时检测5-甲基胞嘧啶和n6 -甲基腺苷的新型光电化学策略

本文首次制备了UiO-66@CdTe@AuNPs复合材料，该复合材料具有光电化学（PEC）传感矩阵的功能，并共轭为三向结（TWJ）。我们提出了一种基于抗体的特异性识别诱导的邻居连接反应，该反应启动两种分子设计的核酸酶（MNAzymes）的组装，释放一种寡核苷酸，该寡核苷酸通过支点介导的链位移反应（TSDR）水解TWJ。随后，形成了一种基于杂化链反应应用（HRCA）的树状大分子，该树状大分子固定了大量量子点，产生了减少光电流信号的突发效应。正如预期的那样，所开发的PEC生物传感器对m6A-RNA和m5C-RNA均表现出优异的分析性能，检测限分别为0.309 fM和6.918 aM。该超灵敏同步PEC生物传感器的成功制备为表观遗传学研究、RNA甲基化相关疾病的生物测定、机制研究和临床诊断提供了新的见解。图形抽象

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Cadmium chloride

来源期刊

Microchimica Acta 化学-分析化学

CiteScore

9.80

自引率

5.30%

发文量

410

审稿时长

2.7 months

期刊介绍： As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.