用于活细胞中 8-oxyguanine DNA 糖基化酶活性灵敏成像的功能性 DNA-Zn2+ 配位纳米球。

IF 5.6 1区 化学 Q1 CHEMISTRY, ANALYTICAL Talanta Pub Date : 2024-12-01 Epub Date: 2024-08-29 DOI:10.1016/j.talanta.2024.126779
Yayun Yang, Nan Zhang, Wei Jiang
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引用次数: 0

摘要

灵敏监测活细胞中人类 8-oxyguanine DNA 糖基化酶(hOGG1)的活性有助于了解其在损伤修复中的功能,并评估其在疾病诊断中的作用。本文提出了一种功能性 DNA-Zn2+ 配位纳米球,用于活细胞中 hOGG1 的灵敏成像。该纳米球是通过熵驱动反应(EDR)-脱氧核糖核酸酶(DNAzyme)系统与Zn2+的配位驱动自组装构建的,其中DNAzyme被设计成分裂结构并组装到EDR系统中。当纳米球进入细胞时,细胞中的磷酸盐和 Zn2+ 之间的竞争性配位导致纳米球解体,释放出 DNA 和一些 Zn2+。释放出的 Zn2+ 成为 DNA 酶的辅助因子。在 hOGG1 的存在下,EDR 完成,同时荧光恢复并生成完整的 DNA 酶。在 Zn2+ 的帮助下,DNA 酶不断裂解底物,产生大量荧光信号,从而实现了对 hOGG1 活性的灵敏成像。该纳米球成功实现了对人宫颈癌细胞(HeLa)、人非小细胞肺癌细胞和人正常结肠上皮细胞中 hOGG1 的灵敏成像,并检测了 HeLa 细胞中 hOGG1 活性的变化。这种纳米球可能会为细胞内 hOGG1 的成像和相关生物医学研究提供一种新的工具。
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Functional DNA-Zn2+ coordination nanospheres for sensitive imaging of 8-oxyguanine DNA glycosylase activity in living cells.

Sensitive monitoring of human 8-oxyguanine DNA glycosylase (hOGG1) activity in living cells is helpful to understand its function in damage repair and evaluate its role in disease diagnosis. Herein, a functional DNA-Zn2+ coordination nanospheres was proposed for sensitive imaging of hOGG1 in living cells. The nanospheres were constructed through the coordination-driven self-assembly of the entropy driven reaction (EDR) -deoxyribozyme (DNAzyme) system with Zn2+, where DNAzyme was designed to split structure and assembled into the EDR system. When the nanospheres entered the cell, the competitive coordination between phosphate in the cell and Zn2+ leaded to the disintegration of the nanospheres, releasing DNA and some Zn2+. The released Zn2+ acted as a cofactor of DNAzyme. In the presence of hOGG1, the EDR was completed, accompanied by fluorescence recovery and the generation of a complete DNAzyme. With the assistance of Zn2+, DNAzyme continuously cleaved substrates to produce plenty of fluorescence signals, thus achieving sensitive imaging of hOGG1 activity. The nanospheres successfully achieved sensitive imaging of hOGG1 in human cervical cancer cells (HeLa), human non-small cell lung cancer cells and human normal colonic epithelial cells, and assayed changes in hOGG1 activity in HeLa cells. This nanospheres may provide a new tool for intracellular hOGG1 imaging and related biomedical studies.

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来源期刊
Talanta
Talanta 化学-分析化学
CiteScore
12.30
自引率
4.90%
发文量
861
审稿时长
29 days
期刊介绍: Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome. Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.
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