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通过包含ZnO@聚多巴胺和DNAzyme探针的纳米系统检测细胞内微小RNA-21用于癌症诊断

Detection of intracellular microRNA-21 for cancer diagnosis by a nanosystem containing a ZnO@polydopamine and DNAzyme probe.

作者信息

Liu Yuanyuan, Wang Ranran, Zhang Fengxia, Ma Yongshan, Jiang Tianyi

机构信息

School of Municipal and Environmental Engineering, Shandong Jianzhu University Jinan 250101 Shandong P. R. China

Yantai Engineering & Technology College Yantai 264006 Shandong P. R. China.

出版信息

RSC Adv. 2024 Apr 26;14(19):13351-13360. doi: 10.1039/d4ra00636d. eCollection 2024 Apr 22.

Abstract

MicroRNAs (miRNAs) are a series of single-stranded non-coding ribonucleic acid (RNA) molecules which associated closely with various human diseases. Efficient strategies for detecting miRNAs are of great significance to cancer diagnosis and prognosis. Here we provide a novel nanosystem that can be applied for the detection of miRNAs. The nanosystem consists of a single-stranded deoxyribonucleic acid (DNA) probe and a probe carrier. The DNA probe was designed based on a deoxyribozyme (DNAzyme) with several necessary functional sequences and two fluorescent dyes labeled at proper sites. The ZnO@polydopamine (ZnO@PDA) nanomaterial serves not only as a probe carrier, but also as a supplier of Zn that can activate the DNAzyme. The DNA probe will undergo a conformation alteration induced by miRNA-21, which then trigger the DNAzyme catalyzed self-cleavage reaction with the assist of Zn provided by ZnO decomposition under weak acid environment. A change of fluorescent color will occur due to the interruption of fluorescence resonance energy transfer between the two fluorescent dyes, and the dissociated miRNA-21 can repeatedly induce the above responses to amplify the fluorescence signal. The feasibility of the whole procedure was demonstrated by various experiments. This nanosystem showed a good selectivity towards miRNA-21, and under the optimal incubation time of 2 hours, a good linear relationship was obtained in a concentration range of 0.01-2.0 nM with a detection limit of 3.8 pM. In detection, an obvious fluorescence color change from red to green can be observed in the presence of miRNA-21. The results proved that this miRNA detection strategy has a broad application prospect in tumor diagnosis and miRNA related biological studies.

摘要

微小RNA(miRNA)是一系列与多种人类疾病密切相关的单链非编码核糖核酸(RNA)分子。高效检测miRNA的策略对癌症诊断和预后具有重要意义。在此,我们提供了一种可用于检测miRNA的新型纳米系统。该纳米系统由单链脱氧核糖核酸(DNA)探针和探针载体组成。DNA探针基于脱氧核酶(DNAzyme)设计,具有几个必要的功能序列,并在适当位置标记了两种荧光染料。ZnO@聚多巴胺(ZnO@PDA)纳米材料不仅作为探针载体,还作为可激活DNAzyme的锌源。DNA探针会因miRNA - 21诱导发生构象改变,然后在弱酸环境下ZnO分解提供的锌的协助下触发DNAzyme催化的自我切割反应。由于两种荧光染料之间的荧光共振能量转移中断,会发生荧光颜色变化,并且解离的miRNA - 21可以反复诱导上述反应以放大荧光信号。各种实验证明了整个过程的可行性。该纳米系统对miRNA - 21表现出良好的选择性,在2小时的最佳孵育时间下,在0.01 - 2.0 nM的浓度范围内获得了良好的线性关系,检测限为3.8 pM。在检测中,在miRNA - 21存在的情况下可以观察到明显的荧光颜色从红色变为绿色。结果证明,这种miRNA检测策略在肿瘤诊断和miRNA相关生物学研究中具有广阔的应用前景。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d571/11047055/42a4c3c94f7c/d4ra00636d-f1.jpg

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