化学
生物物理学
共焦显微镜
共焦
DNA
细胞内
小RNA
荧光显微镜
多重位移放大
聚合
荧光
荧光寿命成像显微镜
核酸
流离失所(心理学)
纳米技术
细胞生物学
生物化学
基因
聚合物
材料科学
生物
光学
聚合酶链反应
有机化学
物理
DNA提取
心理治疗师
心理学
作者
Zhe Yang,Songbai Zhang,Hui Zhao,Huimin Niu,Zai-Sheng Wu,Huan-Tsung Chang
出处
期刊:Analytical Chemistry
[American Chemical Society]
日期:2018-10-31
卷期号:90 (23): 13891-13899
被引量:33
标识
DOI:10.1021/acs.analchem.8b03063
摘要
The microRNA profiles within living cells are informative for diagnosis and prognosis of human cancers. In the present work, we developed a new sensing strategy based on branched DNA junction-enhanced isothermal circular strand displacement polymerization (B-ICSDP) for the detection and intracellular imaging of microRNAs in living cells of interest. A circular DNA template consisting of three repetitive fragments serves as the scaffold for the self-assembly of sophisticated signaling probes, resulting a shrunk branched DNA junction. Target microRNA triggers the opening of molecular beacon, not only restoring the quenched fluorescence but also activating a circular polymerization-based strand displacement reaction. Thus, patulous branched DNA junction is abundantly formed, generating the amplified signal. It is noteworthy that great heaps of branched product assemblies can be also achieved in living cells, and the intracellular enzymatic assembly based strategy is able to be used to recognize specific microRNA-expressed cancer cells. Moreover, different microRNAs coexisting in the same living cells can be simultaneously screened without any interference from each other by confocal laser scanning microscopy. The measured data from confocal fluorescence imaging of different cancer cells demonstrates that the B-ICSDP-based system is a promising alternative for in vivo analysis of microRNAs in complicated biological samples.
科研通智能强力驱动
Strongly Powered by AbleSci AI