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一种光响应和金属有机框架封装的 DNA 四面体熵驱动放大器,用于高性能成像细胞内 microRNA。

A Photoresponsive and Metal-Organic Framework Encapsulated DNA Tetrahedral Entropy-Driven Amplifier for High-Performance Imaging Intracellular MicroRNA.

机构信息

Hubei Province Key Laboratory of Occupational Hazard Identification and Control, School of Public Health, Medical College, Wuhan University of Science and Technology, Wuhan 430065, People's Republic of China.

College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, People's Republic of China.

出版信息

Anal Chem. 2021 Dec 14;93(49):16638-16645. doi: 10.1021/acs.analchem.1c04105. Epub 2021 Dec 2.

DOI:10.1021/acs.analchem.1c04105
PMID:34855353
Abstract

The further development of high-performance fluorescent biosensors to image intracellular microRNAs is beneficial to cancer medicine. By virtue of the need for enzymes and hairpin DNA probes, the entropy-driven reaction-assisted signal amplification strategy has shown an enormous potential to accomplish this task. Nevertheless, this good option still meets with poor biostability, low cell uptake efficiency, and unsatisfactory accuracy. On the basis of these challenges, we put forward here a battery of solving pathways. First, the straight DNA probes are anchored onto the vertexes of dual DNA tetrahedrons, and thus the enzyme resistance of the whole sensing system is observably enhanced. A metal-organic framework (ZIF-8 nanoparticle), which can be effectively dissociated into a weakly acidic environment, then is employed as an additional delivery vehicle to encapsulate such a DNA tetrahedron sustained biosensor and finally bring about a more efficient endocytosis. Last, a kind of photocleavage-linker triggered photoresponsive manner is incorporated to achieve an exceptional precise target identification, by which the biosensor can only be initiated under the irradiation of an externally mild 365 nm ultraviolet light source. In accordance with the above efforts, worthy assay performance toward microRNA-196a has given rise to this newly constructed biosensor, whose sensitivity is down to 2.7 pM and also able to distinguish single-base variation. Beyond that, the amplifier can work as a powerful imaging toolbox to accurately determine the targets in living cells, providing a promising intracellular sensing platform.

摘要

进一步开发用于对细胞内 microRNA 成像的高性能荧光生物传感器有利于癌症医学。由于需要酶和发夹 DNA 探针,熵驱动反应辅助信号放大策略在完成这项任务方面显示出了巨大的潜力。然而,这种好的选择仍然存在生物稳定性差、细胞摄取效率低和准确性不令人满意的问题。针对这些挑战,我们提出了一系列解决方案。首先,将直链 DNA 探针固定在双 DNA 四面体的顶点上,从而显著提高了整个传感系统的酶抗性。然后,将可以有效在弱酸环境中解离的金属有机框架(ZIF-8 纳米颗粒)用作额外的递送载体,封装这种 DNA 四面体持续型生物传感器,最终实现更有效的内吞作用。最后,引入一种光裂解连接体触发的光响应方式,实现了异常精确的目标识别,只有在外部温和的 365nm 紫外光源照射下,生物传感器才能被启动。通过以上努力,对 microRNA-196a 的有价值的检测性能为新构建的生物传感器提供了支持,其灵敏度低至 2.7 pM,并且能够区分单碱基变异。此外,该放大器可以作为一种强大的成像工具包,准确地确定活细胞中的靶标,为细胞内传感提供了一个有前途的平台。

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