In vitro quantification of specific microRNA using molecular beacons.
In vitro quantification of specific microRNA using molecular beacons.
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DOI：
10.1093/nar/gkr1016
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发表时间：
2012-01
影响因子：
14.9
通讯作者：
Searles CD
中科院分区：
文献类型：
--
作者：
Baker MB;Bao G;Searles CD
MicroRNAs （miRNAs）是一类非编码rna，由于其不同的基因组起源和调控一系列细胞过程的能力，已成为分子生物学研究的主要焦点。尽管miRNA的生物学功能尚不完全清楚，但特定miRNA的组织水平已被证明与疾病的病理发展相关。在这里，我们证明了分子信标可以很容易地区分成熟和pre-miRNA，并可靠地量化miRNA的表达。我们发现，DNA、RNA和结合锁定核酸(LNA) -DNA骨架的分子信标在体外均能检测到低浓度（<1 nM）的mirna，其中RNA信标的检测灵敏度最高。此外，我们发现分子信标具有区分核苷酸序列略有变化的mirna的潜力。这些结果表明，基于分子信标的方法评估miRNA表达并区分成熟和前体miRNA物种是非常可靠的，并且有望评估生物样品中的miRNA水平。
MicroRNAs (miRNAs), a class of non-coding RNAs, have become a major focus of molecular biology research because of their diverse genomic origin and ability to regulate an array of cellular processes. Although the biological functions of miRNA are yet to be fully understood, tissue levels of specific miRNAs have been shown to correlate with pathological development of disease. Here, we demonstrate that molecular beacons can readily distinguish mature- and pre-miRNAs, and reliably quantify miRNA expression. We found that molecular beacons with DNA, RNA and combined locked nucleic acid (LNA)–DNA backbones can all detect miRNAs of low (<1 nM) concentrations in vitro, with RNA beacons having the highest detection sensitivity. Furthermore, we found that molecular beacons have the potential to distinguish miRNAs that have slight variations in their nucleotide sequence. These results suggest that the molecular beacon-based approach to assess miRNA expression and distinguish mature and precursor miRNA species is quite robust, and has the promise for assessing miRNA levels in biological samples.
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