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基于 DNA 酶的纳米机器用于活细胞中 microRNA 的原位检测。

DNAzyme Based Nanomachine for in Situ Detection of MicroRNA in Living Cells.

机构信息

Shandong Provincial Key Laboratory of Detection Technology for Tumor Markers, College of Chemistry and Chemical Engineering, Linyi University , Linyi 276005, P. R. China.

Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences, Deakin University , Geelong, Victoria 3217, Australia.

出版信息

ACS Sens. 2017 Dec 22;2(12):1847-1853. doi: 10.1021/acssensors.7b00710. Epub 2017 Dec 12.

Abstract

The capability of in situ detection of microRNA in living cells with signal amplification strategy is of fundamental importance, and it will open up a new opportunity in development of diagnosis and prognosis of many diseases. Herein we report a swing DNA nanomachine for intracellular microRNA detection. The surfaces of Au nanoparticles (NPs) are modified by two hairpin DNA. We observe that one DNA (MB2) will open its hairpin structure upon partial hybridization with target miR-21 after entering into cells, and the other part of its hairpin structure could further react with the other hairpin DNA (MB1) to form a Zn-specific DNAzyme. This results in the disruption of MB1 through shearing action and the release of fluorescein Cy5. To provide an intelligent DNA nanomachine, MB2 is available again with the shearing action to bind with MB1, which provides effective signal amplification. This target-responsive, DNA nanomachine-based method showed a detection limit of 0.1 nM in vitro, and this approach could be an important step toward intracellular amplified detection and imaging of various analytes in living cells.

摘要

利用信号放大策略实现活细胞中 microRNA 的原位检测能力具有重要意义,它将为许多疾病的诊断和预后开辟新的机会。本文报道了一种用于细胞内 microRNA 检测的摆动 DNA 纳米机器。金纳米粒子(AuNPs)的表面修饰有两条发夹 DNA。我们观察到,当进入细胞后,其中一条 DNA(MB2)与靶标 miR-21 部分杂交后会打开其发夹结构,其发夹结构的另一部分可以进一步与另一条发夹 DNA(MB1)反应形成 Zn 特异性 DNA 酶。这导致 MB1 通过剪切作用被破坏,并释放出荧光素 Cy5。为了提供智能 DNA 纳米机器,MB2 可再次通过剪切作用与 MB1 结合,从而提供有效的信号放大。这种基于靶标响应的 DNA 纳米机器在体外的检测限为 0.1 nM,这一方法可能是在活细胞中对各种分析物进行细胞内放大检测和成像的重要一步。

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