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利用 AgNC@NA 的荧光开关和富含鸟嘌呤的突出序列检测癌症相关 miRNA。

Detection of cancer-associated miRNA using a fluorescence switch of AgNC@NA and guanine-rich overhang sequences.

机构信息

Department of Physics, University of Nebraska Omaha, 6001 Dodge Street, Omaha, Nebraska, USA.

出版信息

Luminescence. 2023 Jul;38(7):1385-1392. doi: 10.1002/bio.4471. Epub 2023 Mar 7.

Abstract

DNA-templated silver nanoclusters (AgNC@DNA) are a novel type of nanomaterial with advantageous optical properties. Only a few atoms in size, the fluorescence of nanoclusters can be tuned using DNA overhangs. In this study, we explored the properties of AgNCs manufactured on a short single-stranded (dC) when adjacent G-rich sequences (dG , with N = 3-15) were added. The 'red' emission of AgNC@dC with λ  = 660 nm dramatically changed upon the addition of a G-rich overhang with N  = 15. The pattern of the emission-excitation matrix (EEM) suggested the emergence of two new emissive states at λ  = 575 nm and λ  = 710 nm. The appearance of these peaks provides an effective way to design biosensors capable of detecting specific nucleic acid sequences with low fluorescence backgrounds. We used this property to construct an NA-based switch that brings AgNC and the G overhang near one another, turning 'ON' the new fluorescence peaks only when a specific miRNA sequence is present. Next, we tested this detection switch on miR-371, which is overexpressed in prostate cancer. The results presented provide evidence that this novel fluorescent switch is both sensitive and specific with a limit of detection close to 22 picomoles of the target miR-371 molecule.

摘要

基于 DNA 的银纳米团簇 (AgNC@DNA) 是一种具有优势光学特性的新型纳米材料。纳米团簇的尺寸只有几个原子大小,可以通过 DNA 突出物来调整其荧光。在这项研究中,我们研究了在短单链 (dC) 上制造的 AgNCs 的特性,当相邻的富含 G 序列 (dG,N=3-15) 被添加时。当添加富含 G 的突出物 (N=15) 时,AgNC@dC 的“红色”发射 (λ=660nm) 会发生明显变化。发射-激发矩阵 (EEM) 的模式表明在 λ=575nm 和 λ=710nm 处出现了两个新的发射态。这些峰的出现为设计能够以低荧光背景检测特定核酸序列的生物传感器提供了一种有效方法。我们利用这一特性构建了一个基于 NA 的开关,使 AgNC 和 G 突出物彼此靠近,只有当存在特定的 miRNA 序列时,才会出现新的荧光峰。接下来,我们在前列腺癌中过度表达的 miR-371 上测试了这种检测开关。结果表明,这种新型荧光开关具有高灵敏度和特异性,其检测目标 miR-371 分子的检测限接近 22 皮摩尔。

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