Miao Peng, Tang Yuguo
Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, P. R. China.
Department of Chemistry, New York University, New York 10003, United States.
Anal Chem. 2020 Sep 15;92(18):12700-12709. doi: 10.1021/acs.analchem.0c03181. Epub 2020 Sep 1.
Messenger RNA (mRNA) is a type of important gene regulation element and reliable biomarker for the early diagnosis of diseases. However, mRNA analysis in live cells cannot be easily realized since conventional techniques always require procedures like mRNA purification or cell fixation. Herein, we propose a powerful two-dimensional hybridization chain reaction (2D HCR) strategy for amplified sensing and imaging of intracellular mRNA. The basis is the design of ternary hairpin or dumbbell-structured DNA fuel strands. In the presence of mRNA, organized 2D DNA assembly reaction can be initiated, which leads to structural changes of a large number of fuel strands and produces an in situ 2D DNA network. An enhanced fluorescence signal is thus generated, allowing direct and quantitative analysis of mRNA with high signal-to-noise ratio. Moreover, MnO nanosheet/glutathione-aided transportation and release association are successfully applied to adapt the 2D HCR nanosystem in live cells. Therefore, this work provides a promising quantitative endogenous mRNA analysis tool for clinical applications.
信使核糖核酸（mRNA）是一种重要的基因调控元件，也是疾病早期诊断的可靠生物标志物。然而，由于传统技术总是需要mRNA纯化或细胞固定等程序，活细胞中的mRNA分析不易实现。在此，我们提出了一种强大的二维杂交链式反应（2D HCR）策略，用于细胞内mRNA的扩增传感和成像。其基础是三元发夹或哑铃结构的DNA燃料链的设计。在mRNA存在的情况下，可以启动有组织的二维DNA组装反应，这会导致大量燃料链的结构变化，并产生原位二维DNA网络。从而产生增强的荧光信号，允许以高信噪比直接定量分析mRNA。此外，二氧化锰纳米片/谷胱甘肽辅助的运输和释放关联成功应用于使二维HCR纳米系统适用于活细胞。因此，这项工作为临床应用提供了一种有前景的定量内源性mRNA分析工具。
