Cissell Kyle A, Deo Sapna K
Department of Chemistry and Chemical Biology, Indiana University Purdue University Indianapolis, Indianapolis, IN 46217, USA.
Anal Bioanal Chem. 2009 Jun;394(4):1109-16. doi: 10.1007/s00216-009-2744-6. Epub 2009 Apr 15.
MicroRNAs (miRNAs) are short, approximately 22 nucleotide length RNAs that perform gene regulation. Recently, miRNA has been shown to be linked with the onset of cancer and other diseases based on miRNA expression levels. It is important, therefore, to understand miRNA function as it pertains to disease onset; however, in order to fully understand miRNA's role in a disease, it is necessary to detect the expression levels of these small molecules. The most widely used miRNA detection method is Northern blotting, which is considered as the standard of miRNA detection methods. This method, however, is time-consuming and has low sensitivity. This has led to an increase in the amount of detection methods available. These detection methods are either solid phase, occurring on a solid support, or solution phase, occurring in solution. While the solid-phase methods are adaptable to high-throughput screening and possess higher sensitivity than Northern blotting, they lack the ability for in vivo use and are often time-consuming. The solution-phase methods are advantageous in that they can be performed in vivo, are very sensitive, and are rapid; however, they cannot be applied in high-throughput settings. Although there are multiple detection methods available, including microarray technology, luminescence-based assays, electrochemical assays, etc., there is still much work to be done regarding miRNA detection. The current gaps of miRNA detection include the ability to perform multiplex, sensitive detection of miRNA with single-nucleotide specificity along with the standardization of these new methods. Current miRNA detection methods, gaps in these methods, miRNA therapeutic options, and the future outlook of miRNA detection are presented here.
微小RNA(miRNA)是一类短链RNA,长度约为22个核苷酸,具有基因调控功能。最近的研究表明,基于miRNA的表达水平,其与癌症及其他疾病的发生有关。因此,了解miRNA在疾病发生中的作用非常重要;然而,为了全面理解miRNA在疾病中的作用,有必要检测这些小分子的表达水平。最常用的miRNA检测方法是Northern印迹法,它被认为是miRNA检测方法的标准。然而,这种方法耗时且灵敏度低。这导致了可用检测方法数量的增加。这些检测方法要么是固相的,即在固体支持物上进行,要么是液相的,即在溶液中进行。虽然固相方法适用于高通量筛选,且比Northern印迹法具有更高的灵敏度,但它们缺乏体内应用的能力,且通常耗时。液相方法的优点在于它们可以在体内进行,非常灵敏且快速;然而,它们不能应用于高通量检测。尽管有多种检测方法可用,包括微阵列技术、基于发光的检测方法、电化学检测方法等,但在miRNA检测方面仍有许多工作要做。miRNA检测目前存在的差距包括能够对miRNA进行具有单核苷酸特异性的多重、灵敏检测以及这些新方法的标准化。本文介绍了当前的miRNA检测方法、这些方法存在的差距、miRNA治疗选择以及miRNA检测的未来展望。