Photoactivated Specific mRNA Detection in Single Living Cells by Coupling “Signal-on” Fluorescence and “Signal-off” Electrochemical Signals,Nano Letters

当前位置： X-MOL 学术 › Nano Lett. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Photoactivated Specific mRNA Detection in Single Living Cells by Coupling “Signal-on” Fluorescence and “Signal-off” Electrochemical Signals
Nano Letters ( IF 9.6 ) Pub Date : 2018-07-12 00:00:00 , DOI: 10.1021/acs.nanolett.8b02004
Fujian Huang ₁ , Meihua Lin ₁ , Ruilin Duan ₁ , Xiaoding Lou ₁ , Fan Xia _{1,

2} , Itamar Willner ₃

Affiliation

The spatiotemporal detection of a target mRNA in a single living cell is a major challenge in nanoscience and nanomedicine. We introduce a versatile method to detect mRNA at a single living cell level that uses photocleavable hairpin probes as functional units for the optical (fluorescent) and electrochemical (voltammetric) detection of MnSOD mRNA in single MCF-7 cancer cells. The fluorescent probe is composed of an ortho-nitrophenylphosphate ester functionalized hairpin that includes the FAM fluorophore in a caged configuration quenched by Dabcyl. The fluorescent probe is further modified with the AS1411 aptamer to facilitate the targeting and internalization of the probe into the MCF-7 cells. Under UV irradiation, the hairpin is cleaved, leading to the intracellular mRNA toehold-stimulated displacement of the FAM-functionalized strand resulting in a switched-on fluorescence signal upon the detection of the mRNA in a single cell. In addition, a nanoelectrode functionalized with a methylene blue (MB) redox-active photocleavable hairpin is inserted into the cytoplasm of a single MCF-7 cell. Photocleavage of the hairpin leads to the mRNA-mediated toehold displacement of the redox-active strand associated with the probe, leading to the depletion of the voltammetric response of the probe. The parallel optical and electrochemical detection of the mRNA at a single cell level is demonstrated.

中文翻译：

通过“信号开启”荧光和“信号关闭”电化学信号的耦合，在单个活细胞中进行光活化特异性mRNA检测。

在单个活细胞中对目标mRNA的时空检测是纳米科学和纳米医学中的主要挑战。我们介绍了一种在单个活细胞水平上检测mRNA的通用方法，该方法使用可光裂解的发夹探针作为功能单元，用于单个MCF-7癌细胞中MnSOD mRNA的光学（荧光）和电化学（伏安）检测。荧光探针由邻硝基苯基磷酸酯官能化的发夹组成，该发夹包括呈笼状构型的FAM荧光团，并用Dabcyl淬灭。荧光探针用AS1411适体进一步修饰，以促进探针靶向和内化到MCF-7细胞中。在紫外线照射下，发夹被劈开，导致FAM功能化链的胞内mRNA脚趾头刺激刺激的位移，导致在单个细胞中检测到mRNA后打开荧光信号。此外，将用亚甲蓝（MB）氧化还原活性的光可裂解发夹功能化的纳米电极插入单个MCF-7细胞的细胞质中。发夹的光裂解导致与探针相关的氧化还原活性链的mRNA介导的脚趾位移，从而导致探针的伏安反应耗竭。证明了在单个细胞水平上mRNA的并行光学和电化学检测。将用亚甲蓝（MB）氧化还原活性的光可裂解发夹功能化的纳米电极插入单个MCF-7细胞的细胞质中。发夹的光裂解导致与探针相关的氧化还原活性链的mRNA介导的脚趾位移，从而导致探针的伏安反应耗竭。证明了在单个细胞水平上mRNA的并行光学和电化学检测。将用亚甲蓝（MB）氧化还原活性的光可裂解发夹功能化的纳米电极插入单个MCF-7细胞的细胞质中。发夹的光裂解导致与探针相关的氧化还原活性链的mRNA介导的脚趾位移，从而导致探针的伏安反应耗竭。证明了在单个细胞水平上mRNA的并行光学和电化学检测。

更新日期：2018-07-12

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11