Fan Yaofang, Liu Yanming, Zhou Qihui, Du Hao, Zhao Xueyang, Ye Fei, Zhao Huimin
Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education, China), School of Environmental Science and Technology, Dalian University of Technology, Dalian, 116024, China.
Institute for Translational Medicine, Department of Stomatology, The Affiliated Hospital of Qingdao University, Qingdao, 266003, China.
Talanta. 2021 Feb 1;223(Pt 1):121675. doi: 10.1016/j.talanta.2020.121675. Epub 2020 Sep 28.
Fluorescence resonance energy transfer, a promising method for in situ imaging of miRNA in living cells, has intrinsic limitation on sensitivity and selectivity. Herein, a fluorescent amplification strategy based on catalyzed hairpin assembly indirectly covalent on FeO@C nanoparticles via short single-stranded DNA was investigated for cellular miRNA detection in living cells, integrating non-enzyme target-active releasing for amplifying the signal output, highly quenching efficiency of FeO@C nanoparticles with low background, ssDNA assisted fluorescent group-fueled chain releasing from FeO@C nanoparticles with enhanced fluorescence response. The designed platform exhibits highly sensitive in a wide linear concentration range of 0.450 pM-190 pM and is highly specific for miRNA-20a detection with the ability of discriminating one mistake base. Additionally, the CHA-FeO@C was successfully applied in imaging visualization of miRNA-20a in the living cell. The strategy provides a promising bioassay approach for clinical research.
荧光共振能量转移是一种用于活细胞中miRNA原位成像的有前景的方法，但在灵敏度和选择性方面存在固有局限性。在此，研究了一种基于催化发夹组装的荧光放大策略，该策略通过短单链DNA间接共价连接在FeO@C纳米颗粒上，用于活细胞中细胞miRNA的检测，整合了非酶靶标活性释放以放大信号输出、FeO@C纳米颗粒的高淬灭效率和低背景、ssDNA辅助荧光基团从FeO@C纳米颗粒释放以增强荧光响应。所设计的平台在0.450 pM-190 pM的宽线性浓度范围内表现出高度敏感性，并且对miRNA-20a检测具有高度特异性，能够区分一个错配碱基。此外，CHA-FeO@C成功应用于活细胞中miRNA-20a的成像可视化。该策略为临床研究提供了一种有前景的生物测定方法。
