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基于集成化和自动化 DNA 行走器的电化学 DNA 传感器。

An electrochemical DNA sensor based on an integrated and automated DNA walker.

机构信息

JiangXi University of Chinese Medicine, Nan Chang, JiangXi 330004, China.

JiangXi University of Chinese Medicine, Nan Chang, JiangXi 330004, China.

出版信息

Bioelectrochemistry. 2022 Oct;147:108198. doi: 10.1016/j.bioelechem.2022.108198. Epub 2022 Jun 28.

Abstract

As an artificial nanomachine, a DNA walker demonstrates the potential for biosensing. In this study, a highly integrated, biostable, and autonomous electrochemical DNA walker sensor was rationally designed by a simple assembly of a Mn-dependent DNAzyme-powered DNA walker with nanoscale Mn @MOFs containing free carboxylic acid groups UiO-66(Zr)-(COOH). In this study, the release of Mn from Mn@MOFs was exploited to drive the autonomous and progressive operation of the DNA walker, and the DNAzyme-driven DNA walker was constructed by the co-modification of walking strands and track strands onto the gold electrode (GE) surface. The walking strand was a single-stranded DNA containing a DNAzyme sequence, which was pre-silenced by the locking strand. The track strand was a specially designed DNA sequence that the target can hybridize with the locking strand; hence, the walking strand is unlocked, and the liberated DNAzyme catalyzes the cleavage of track strands to drive the DNA walker operation, shifting tetraferrocene away from the electrode and producing a significant signal change. A detection limit of 38 fM was obtained with our new system, exhibiting a wide linear range from 1.5625 × 10 M to 1 × 10 M. The proposed approach provided a novel means for constructing an highly integrated, automated, and DNAzyme-driven DNA walker for bioanalysis.

摘要

作为一种人工纳米机器,DNA walker 展示了用于生物传感的潜力。在这项研究中,通过将 Mn 依赖性 DNA 酶驱动的 DNA walker 与具有游离羧酸基团 UiO-66(Zr)-(COOH)的纳米级 Mn@MOFs 进行简单组装,合理设计了一种高度集成、生物稳定和自主的电化学 DNA walker 传感器。在这项研究中,利用 Mn@MOFs 中 Mn 的释放来驱动 DNA walker 的自主和渐进式操作,并且通过将行走链和轨道链共修饰到金电极(GE)表面来构建 DNA 酶驱动的 DNA walker。行走链是一条单链 DNA,其中包含一个 DNA 酶序列,该序列被锁定链预先沉默。轨道链是一条经过特殊设计的 DNA 序列,其可以与锁定链杂交;因此,行走链被解锁,并且释放的 DNA 酶可以催化轨道链的切割以驱动 DNA walker 操作,从而将四茂铁从电极上转移并产生显著的信号变化。我们的新系统获得了 38 fM 的检测限,表现出从 1.5625×10-15 M 到 1×10-13 M 的宽线性范围。该方法为构建用于生物分析的高度集成、自动化和 DNA 酶驱动的 DNA walker 提供了一种新方法。

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