Single Cell Real-Time miRNAs Sensing Based on Nanomotors
Single Cell Real-Time miRNAs Sensing Based on Nanomotors
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DOI：
10.1021/acsnano.5b02807
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发表时间：
2015-07-01
期刊：
影响因子：
17.1
通讯作者：
Wang, Joseph
中科院分区：
文献类型：
--
作者：
de Avila, Berta Esteban-Fernandez;Martin, Aida;Wang, Joseph
描述了一种基于纳米马达的快速单步细胞内生物传感靶miRNA的策略，靶miRNA在完整癌细胞中表达，在单细胞水平上。这个新概念依赖于超声波（US）推进染料标记单链DNA (ssDNA)/氧化石墨烯（GO）涂层金纳米线（AuNWs）的使用，这些纳米线能够穿透完整的癌细胞。一旦纳米马达被内化到细胞中，猝灭的荧光信号（由氧化石墨烯和染料标记的ssDNA之间的pi-pi相互作用产生）就会被恢复，这是由于染料-ssDNA探针在与靶miRNA-21结合时从马达的氧化石墨烯猝灭表面位移，导致细胞内的“OFF-ON”荧光开关。美国动力纳米马达更快的内化过程及其快速进入细胞的运动增加了探针目标接触的可能性，从而实现了高效快速的杂交。通过测量miRNA-21表达水平存在显著差异的两种类型的癌细胞（MCF-7和HeLa）的荧光信号，证明了基于纳米马达的方法基于目标miRNA内源性含量筛选癌细胞的能力。这种单步、基于运动的miRNA传感方法能够快速“移动”特异性检测目标miRNA-21，即使在内源性miRNA-21含量极低的单细胞中，也可以精确实时地监测细胞内miRNA表达。
A nanomotor-based strategy for rapid single-step intracellular biosensing of a target miRNA, expressed in intact cancer cells, at the single cell level is described. The new concept relies on the use of ultrasound (US) propelled dye-labeled single-stranded DNA (ssDNA)/graphene-oxide (GO) coated gold nanowires (AuNWs) capable of penetrating intact cancer cells. Once the nanomotor is internalized into the cell, the quenched fluorescence signal (produced by the pi-pi interaction between GO and a dye-labeled ssDNA) is recovered due to the displacement of the dye-ssDNA probe from the motor GO-quenching surface upon binding with the target miRNA-21, leading to an attractive intracellular "OFF-ON" fluorescence switching. The faster internalization process of the US-powered nanomotors and their rapid movement into the cells increase the likelihood of probe target contacts, leading to a highly efficient and rapid hybridization. The ability of the nanomotor-based method to screen cancer cells based on the endogenous content of the target miRNA has been demonstrated by measuring the fluorescence signal in two types of cancer cells (MCF-7 and HeLa) with significantly different miRNA-21 expression levels. This single-step, motor-based miRNAs sensing approach enables rapid "on the move" specific detection of the target miRNA-21, even in single cells with an extremely low endogenous miRNA-21 content, allowing precise and real-time monitoring of intracellular miRNA expression.