Liu Yuanyuan, Wang Ranran, Zhang Fengxia, Ma Yongshan, Jiang Tianyi
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.
RSC Adv. 2024 Apr 26;14(19):13351-13360. doi: 10.1039/d4ra00636d. eCollection 2024 Apr 22.
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.
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。