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用于检测微小RNA的荧光共振能量转移(FRET)生物传感器膜的制备。

Production of a fluorescence resonance energy transfer (FRET) biosensor membrane for microRNA detection.

作者信息

Fu Yike, Chen Tong, Wang Gang, Gu Tongxu, Xie Congkun, Huang Jie, Li Xiang, Best Serena, Han Gaorong

机构信息

State Key Laboratory of Silicon Materials, School of Materials Science and Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, P. R. China.

出版信息

J Mater Chem B. 2017 Sep 14;5(34):7133-7139. doi: 10.1039/c7tb01399j. Epub 2017 Aug 17.

Abstract

MicroRNAs (miRNAs) play a key role in regulating gene expression but can be associated with abnormalities linked to carcinogenesis and tumor progression. Hence there is increasing interest in developing methods to detect these non-coding RNA molecules in the human circulation system. Here, a novel FRET miRNA-195 targeting biosensor, based on silica nanofibers incorporated with rare earth-doped calcium fluoride particles (CaF:Yb,Ho@SiO) and gold nanoparticles (AuNPs), is reported. The formation of a sandwich structure, as a result of co-hybridization of the target miRNA which is captured by oligonucleotides conjugated at the surface of CaF:Yb,Ho@SiO fibers and AuNPs, brings the nanofibers and AuNPs in close proximity and triggers the FRET effect. The intensity ratio of green to red emission, I/I, was found to decrease linearly upon increasing the concentration of the target miRNA and this can be utilized as a standard curve for quantitative determination of miRNA concentration. This assay offers a simple and convenient method for miRNA quantification, with the potential for rapid and early clinical diagnosis of diseases such as breast cancer.

摘要

微小RNA(miRNA)在调节基因表达中起关键作用,但可能与致癌作用和肿瘤进展相关的异常有关。因此,人们越来越关注开发在人体循环系统中检测这些非编码RNA分子的方法。在此,报道了一种基于掺入稀土掺杂氟化钙颗粒(CaF:Yb,Ho@SiO)和金纳米颗粒(AuNP)的二氧化硅纳米纤维的新型FRET miRNA-195靶向生物传感器。由于靶标miRNA的共杂交形成了夹心结构,该靶标miRNA被缀合在CaF:Yb,Ho@SiO纤维和AuNP表面的寡核苷酸捕获,使纳米纤维和AuNP紧密靠近并触发FRET效应。发现随着靶标miRNA浓度的增加,绿色与红色发射的强度比I/I呈线性下降,这可作为定量测定miRNA浓度的标准曲线。该测定法为miRNA定量提供了一种简单方便的方法,具有对乳腺癌等疾病进行快速早期临床诊断的潜力。

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