Yin Viravuth P
Mount Desert Island Biological Laboratory, Kathryn W. Davis Center for Regenerative Biology and Medicine, P.O. Box 35, Salisbury Cove, ME, 04672, USA.
Methods Mol Biol. 2018;1649:197-208. doi: 10.1007/978-1-4939-7213-5_13.
Elucidating the spatial resolution of gene transcripts provides important insight into potential gene function. MicroRNAs are short, singled-stranded noncoding RNAs that control gene expression through base-pair complementarity with target mRNAs in the 3' untranslated region (UTR) and inhibiting protein expression. However, given their small size of ~22- to 24-nt and low expression levels, standard in situ hybridization detection methods are not amendable for microRNA spatial resolution. Here, I describe a technique that employs RNAscope probe design and propriety amplification technology that provides simultaneous single molecule detection of individual microRNA and its target gene. This method allows for rapid and sensitive detection of noncoding RNA transcripts in frozen tissue sections.
阐明基因转录本的空间分辨率有助于深入了解潜在的基因功能。微小RNA是短的单链非编码RNA，通过与3'非翻译区（UTR）中的靶mRNA碱基对互补来控制基因表达，并抑制蛋白质表达。然而，由于其大小约为22至24个核苷酸且表达水平较低，标准的原位杂交检测方法不适用于微小RNA的空间分辨率检测。在此，我描述了一种技术，该技术采用RNAscope探针设计和适当的扩增技术，可同时对单个微小RNA及其靶基因进行单分子检测。该方法能够快速、灵敏地检测冰冻组织切片中的非编码RNA转录本。
