劈理(地质)
核糖核酸
滚动圆复制
小RNA
计算生物学
化学
生物
分子生物学
基因
细胞生物学
生物化学
聚合酶
断裂(地质)
古生物学
作者
Marlen Hesse,Christoph Arenz
出处
期刊:ChemBioChem
[Wiley]
日期:2015-12-17
卷期号:17 (4): 304-307
被引量:7
标识
DOI:10.1002/cbic.201500537
摘要
Micro RNA (miRNA) research has evolved into an essential part of investigating gene regulation in which deregulation of numerous miRNAs is associated with various cellular dysfunction and diseases. Here, we describe a rapid and homogenous assay for Ago2-mediated target RNA cleavage, based on branched rolling circle amplification (BRCA). In particular, the ability to investigate small molecule binders for inhibition of miRNA function is within the potential of our assay. This method uses no artificial fluorescence labeling of RNA components, which can be an advantage in screening of potential inhibitors. To visualize cleavage of RNA substrate by Ago2, we developed a two-step assay composed of Ago2-mediated cleavage and BRCA-based detection. The assay is cost-effective and practicable and can be performed in 96-well format by using a standard qPCR machine.
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