Construction of an efficient microRNA sensing platform based on terminal deoxynucleotidyl transferase-mediated synthesis of copper nanoclusters

IF 8 1区化学 Q1 CHEMISTRY, ANALYTICAL

Sensors and Actuators B: Chemical Pub Date : 2024-11-04 DOI:10.1016/j.snb.2024.136892

Yujuan Qi, Yan Chen, Yun Huang, Baoting Dou, Sen Zheng, Zhenbei Li, Po Wang, Shengkun Zhang

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引用次数: 0

Abstract

A conceptual innovative electrochemical sensing platform was constructed for microRNA (miRNA) detection based on terminal deoxynucleotidyl transferase (TdT)-mediated synthesis of copper nanoclusters. In principle, the substrate strand immobilized on working electrode acted as the template for the TdT-mediated DNA extension reaction, while the DNAzyme probe would be activated in the absence of miRNA 21, which led to the cleavage of the substrate strand to produce the 5’ phosphate terminus and inhibited the TdT-mediated DNA extension reaction. On the other hand, the presence of miRNA 21 initiated the toehold-mediated strand displacement reaction, causing the disassembly of the DNAzyme structure and preventing the substrate strand from being cleaved. This enabled the recycle amplification of target miRNA 21, and TdT catalyzed the DNA extension reaction on the 3’ hydroxyl terminus of the substrate strand, producing a large number of poly thymine (polyT) sequences. These sequences bound with copper ions for in-situ synthesis of copper nanoclusters on the sensing electrode, and the increased electrochemical signal of copper was recorded by differential pulse stripping voltammetry (DPSV), which paved the way for sensitive determination of miRNA 21 extracted from cancer cells with a detection limit of 36 aM. Due to its high sensitivity, user-friendly operation, and cost-effectiveness, this method is helpful for the fundamental research of sensing mechanism as well as the diagnosis of related diseases.

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基于末端脱氧核苷酸转移酶介导的纳米铜簇合成构建高效 microRNA 传感平台

基于末端脱氧核苷酸转移酶（TdT）介导的纳米铜簇合成，构建了一个用于检测微RNA（miRNA）的概念性创新电化学传感平台。原则上，固定在工作电极上的底物链是 TdT 介导的 DNA 延伸反应的模板，而 DNA 酶探针会在 miRNA 21 缺失的情况下被激活，导致底物链裂解产生 5' 磷酸末端，从而抑制 TdT 介导的 DNA 延伸反应。另一方面，miRNA 21 的存在启动了趾hold 介导的链置换反应，导致 DNA 酶结构解体，阻止了底物链的裂解。这使得目标 miRNA 21 得以循环扩增，TdT 在底物链的 3'羟基末端催化 DNA 延伸反应，产生大量的聚胸腺嘧啶（polyT）序列。这些序列与铜离子结合，在传感电极上原位合成纳米铜团簇，通过差分脉冲剥离伏安法（DPSV）记录铜电化学信号的增加，为灵敏测定从癌细胞中提取的 miRNA 21 铺平了道路，其检测限为 36 aM。该方法灵敏度高、操作简便、成本低廉，有助于传感机制的基础研究和相关疾病的诊断。

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来源期刊

Sensors and Actuators B: Chemical 工程技术-电化学

CiteScore

14.60

自引率

11.90%

发文量

1776

审稿时长

3.2 months

期刊介绍： Sensors & Actuators, B: Chemical is an international journal focused on the research and development of chemical transducers. It covers chemical sensors and biosensors, chemical actuators, and analytical microsystems. The journal is interdisciplinary, aiming to publish original works showcasing substantial advancements beyond the current state of the art in these fields, with practical applicability to solving meaningful analytical problems. Review articles are accepted by invitation from an Editor of the journal.