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Near-infrared triggered strand displacement amplification for MicroRNA quantitative detection in single living cells

机译:用于单个活细胞中MicroRNA定量检测的近红外触发链置换扩增

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摘要

As an important modulator of gene expression, microRNA (miRNA) has been described as a promising biomarker for the early diagnosis of cancers. A non-invasive method for real-time sensitive imaging and monitoring of miRNA in living cells is in urgent demand. Although some amplified methods have been developed, few can be programmed to assemble single intelligent nanostructures to realize sensitive intracellular miRNA detection without extra addition of an enzyme or catalytic fuel. Herein, two programmable oligonucleotide hairpin probe functionalized gold nanorods (THP-AuNRs) were designed to develop a near-infrared (NIR) laser triggered target strand displacement amplification (SDA) approach for sensitive miRNA imaging quantitative analysis in single living cells and multicellular tumor spheroids (MCTSs). Such a NIR-triggered SDA strategy achieves facile and sensitive monitoring of a model oncogenic miRNA-373 in various cancer lines and MCTS simulated tumor tissue. Notably, using a linear regression equation derived from miRNA mimics, a quantitative method of miRNA in single living cells was realized due to the high sensitivity. This provides a new way for sensitive real-time monitoring of intracellular miRNA, and may be promising for miRNA-based biomedical applications.
机译:作为基因表达的重要调节剂,microRNA(miRNA)被描述为癌症早期诊断的有前途的生物标志物。迫切需要一种非侵入性方法,用于实时敏感成像和监测活细胞中的miRNA。尽管已经开发了一些扩增方法,但是几乎不需要编程就可以组装单个智能纳米结构来实现敏感的细胞内miRNA检测,而无需额外添加酶或催化燃料。本文中,设计了两个可编程的寡核苷酸发夹探针功能化的金纳米棒(THP-AuNRs),以开发近红外(NIR)激光触发的靶链置换扩增(SDA)方法,用于在单个活细胞和多细胞肿瘤球体内进行敏感的miRNA成像定量分析(MCTS)。这种NIR触发的SDA策略可在各种癌症细胞系和MCTS模拟的肿瘤组织中实现对模型致癌miRNA-373的简便而灵敏的监测。值得注意的是,使用源自miRNA模拟物的线性回归方程式,由于具有高灵敏度,因此实现了在单个活细胞中定量miRNA的方法。这提供了一种敏感的实时监测细胞内miRNA的新方法,对于基于miRNA的生物医学应用可能很有希望。

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