Ratiometric and amplified fluorescence nanosensor based on a DNA tetrahedron for miRNA imaging in living cells,Journal of Materials Chemistry B

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Ratiometric and amplified fluorescence nanosensor based on a DNA tetrahedron for miRNA imaging in living cells
Journal of Materials Chemistry B ( IF 6.1 ) Pub Date : 2021-08-24 , DOI: 10.1039/d1tb01537k
Liuting Mo ₁ , Danlian Liang ₁ , Wanqi He ₁ , Chan Yang ₁ , Weiying Lin ₁

Affiliation

Enzyme-free signal amplification approaches have attracted considerable attention in the field of intracellular miRNA analysis. However, the application of nucleic acid amplification has been limited by intracellular delivery of multiple oligonucleotide components with precise stoichiometry. In this work, we propose a new DNA tetrahedron (DTN)-based sensing platform addressing the delivery and stoichiometric control of nucleic components for enzyme-free amplification. The nanosensor is composed of two DTN probes; DTN-F served as the target recognition and signal output unit, and DTN-H served as the signal amplification unit. DTNs could facilitate the cell internalization of the nucleic acid probes and protect them from nuclease degradation. In the absence of target miRNA, the fluorescent strands (F) hybridize with the hanging sequences of DTN, and FAM and TAMRA labeled on F will be separated, blocking fluorescence resonance energy transfer (FRET). In the presence of the target miRNA, F will be displaced by the target and the hairpin structure will be restored, bringing the FRET pair into close proximity and inducing a FRET signal. Moreover, the helper strands (H) on DTN-H could liberate target miRNA through strand displacement, which will initiate a new round of reaction, generating an amplified FRET signal. The DTN nanosensor realized sensitive and selective detection of let-7a in buffer solution and 10% FBS solution. In addition, imaging of miRNA in the different cell lines and monitoring of intracellular miRNA fluctuations were carried out The developed method offers a new tool for bioanalytical and biomedical research.

中文翻译：

基于DNA四面体的比例放大荧光纳米传感器用于活细胞中的miRNA成像

无酶信号放大方法在细胞内 miRNA 分析领域引起了相当大的关注。然而，核酸扩增的应用受到具有精确化学计量的多个寡核苷酸组分的细胞内递送的限制。在这项工作中，我们提出了一种新的基于 DNA 四面体 ( DTN ) 的传感平台，用于解决无酶扩增的核酸成分的传递和化学计量控制。纳米传感器由两个DTN探针组成；DTN-F作为目标识别和信号输出单元，DTN-H作为信号放大单元。DTN可以促进核酸探针的细胞内化并保护它们免受核酸酶降解。在没有靶miRNA的情况下，荧光链（F ）与DTN的悬挂序列杂交，F上标记的FAM和TAMRA将分离，阻断荧光共振能量转移（FRET）。在存在目标 miRNA 的情况下，F将被目标置换，发夹结构将恢复，使 FRET 对靠近并诱导 FRET 信号。此外， DTN-H上的辅助链 ( H )可以通过链置换释放靶miRNA，从而启动新一轮反应，产生放大的FRET信号。DTN纳米传感器实现了对缓冲溶液和10% FBS溶液中let-7a的灵敏选择性检测。此外，还对不同细胞系中的 miRNA 进行了成像，并监测了细胞内 miRNA 的波动。所开发的方法为生物分析和生物医学研究提供了新的工具。

更新日期：2021-09-16

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