分子信标
阿尔戈瑙特
小RNA
基因敲除
劈理(地质)
细胞生物学
化学
核酸酶
信标
生物
分子生物学
RNA干扰
核糖核酸
DNA
寡核苷酸
生物化学
基因
计算机科学
古生物学
实时计算
断裂(地质)
作者
Benjamin M. Luby,Gang Zheng
标识
DOI:10.1002/anie.201707366
摘要
MicroRNA detection is a valuable method for determining cell identity. Molecular beacons are elegant sensors that can transform intracellular microRNA concentration into a fluorescence intensity. While target binding enhances beacon fluorescence, the degree of enhancement is insufficient for demanding applications. The addition of specialty nucleases can enable target recycling and signal amplification, but this process complicates the assay. We have developed and characterized a class of beacons that are susceptible to the endogenous nuclease Argonaute-2 (Ago2). After purification of the complex by co-immunoprecipitation, microRNA:Ago2 cleavage (miRACle) beacons undergo site- and sequence-specific cleavage, and show a 13-fold fluorescence enhancement over traditional beacons. The system can be adapted to any microRNA sequence, and can cleave nuclease-resistant, non-RNA bases, potentially allowing miRACle beacons to be designed for cells without interference from non-specific nucleases.
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