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高度集成的 DNA 纳米机器中的 DNA 酶级联电路,用于活细胞中敏感 microRNAs 的成像。

DNAzyme cascade circuits in highly integrated DNA nanomachines for sensitive microRNAs imaging in living cells.

机构信息

State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, PR China.

Department of Biochemistry and Molecular Biology, Key Laboratory of Longevity and Aging-related Diseases of Chinese Ministry of Education, Center for Translational Medicine & School of Basic Medical Sciences, Guangxi Medical University, Nanning, 530021, PR China.

出版信息

Biosens Bioelectron. 2021 Apr 1;177:112976. doi: 10.1016/j.bios.2021.112976. Epub 2021 Jan 6.

Abstract

DNA molecular probes have emerged as powerful tools for fluorescence imaging of microRNAs (miRNAs) in living cells and thus elucidating functions and dynamics of miRNAs. In particular, the highly integrated DNA probes that can be able to address the robustness, sensitivity and consistency issues in a single assay system were highly desired but remained largely unsolved challenge. Herein, we reported for the first time that the development of the novel DNA nanomachines that split-DNAzyme motif was highly integrated in a single DNA triangular prism (DTP) reactor and can undergo target-activated DNAzyme catalytic cascade circuits, allowing amplified sensing and imaging of tumor-related microRNA-21 (miR-21) in living cells. The DNA nanomachines have shown dynamic responses for target miR-21 with excellent sensitivity and selectivity and demonstrated the potential for living cell imaging of miR-21. With the advantages of facile modular design and assembly, high biostability, low cytotoxicity and excellent cellular internalization, the highly integrated DNA nanomachines enabled accurate and effective monitoring of miR-21 expression levels in living cells. Therefore, our developed strategy may afford a reliable and robust nanoplatform for tumor diagnosis and for related biological research.

摘要

DNA 分子探针已成为在活细胞中荧光成像 microRNAs(miRNAs)的有力工具,从而阐明 miRNAs 的功能和动态。特别是,高度集成的 DNA 探针能够在单个检测系统中解决稳健性、灵敏度和一致性问题,这是人们高度期望的,但仍然是一个未解决的主要挑战。在此,我们首次报道了一种新型 DNA 纳米机器的开发,该纳米机器将分裂 DNA 酶基序高度集成在单个 DNA 三角棱柱(DTP)反应器中,并能进行靶标激活的 DNA 酶催化级联反应,从而允许对肿瘤相关 microRNA-21(miR-21)进行放大检测和在活细胞中的成像。DNA 纳米机器对靶标 miR-21 表现出动态响应,具有出色的灵敏度和选择性,并展示了对 miR-21 进行活细胞成像的潜力。由于具有易于模块化设计和组装、高生物稳定性、低细胞毒性和优异的细胞内化能力等优点,高度集成的 DNA 纳米机器能够准确有效地监测活细胞中 miR-21 的表达水平。因此,我们开发的策略可能为肿瘤诊断和相关生物学研究提供一个可靠和稳健的纳米平台。

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