A Target-Triggered DNAzyme Motor Enabling Homogeneous, Amplified Detection of Proteins
A Target-Triggered DNAzyme Motor Enabling Homogeneous, Amplified Detection of Proteins
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DOI：
10.1021/acs.analchem.7b03529
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发表时间：
2017-12-05
影响因子：
7.4
通讯作者：
Zhang, Hongquan
中科院分区：
文献类型：
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作者：
Chen, Junbo;Zuehlke, Albert;Zhang, Hongquan
我们在这里报告的概念，一个自供电，目标触发的DNA马达构建的工程DNA酶，以适应结合诱导的DNA组装。通过DNA间隔区将亲和配体连接到DNAzyme马达，并且将第二亲和配体缀合到金纳米颗粒（AuNP），金纳米颗粒（AuNP）也用数百个底物链装饰，作为DNAzyme马达的高密度三维轨道。靶分子与两种配体的结合诱导DNA酶与其在AuNP上的底物之间的杂交，否则它们不能自发杂交。DNA酶与底物的杂交启动了底物的切割以及DNA酶沿着AuNP的自主运动。每个移动步骤恢复染料分子的荧光，从而能够真实的实时监测DNAzyme马达的操作。辅助因子Mg 2+的简单添加或耗尽允许对DNAzyme马达的精细控制。马达可以将单个结合事件转化为数百种底物的切割，从而能够在室温下放大检测蛋白质，而无需分离。
We report here the concept of a self-powered, target-triggered DNA motor constructed by engineering a DNAzyme to adapt into binding-induced DNA assembly. An affinity ligand was attached to the DNAzyme motor via a DNA spacer, and a second affinity ligand was conjugated to the gold nanoparticle (AuNP) that was also decorated with hundreds of substrate strands serving as a high-density, three-dimensional track for the DNAzyme motor. Binding of a target molecule to the two ligands induced hybridization between the DNAzyme and its substrate on the AuNP, which are otherwise unable to spontaneously hybridize. The hybridization of DNAzyme with the substrate initiates the cleavage of the substrate and the autonomous movement of the DNAzyme along the AuNP. Each moving step restores the fluorescence of a dye molecule, enabling monitoring of the operation of the DNAzyme motor in real time. A simple addition or depletion of the cofactor Mg2+ allows for fine control of the DNAzyme motor. The motor can translate a single binding event into cleavage of hundreds of substrates, enabling amplified detection of proteins at room temperature without the need for separation.