Beckman William, Vuist Ilona M, Kempe Hermannus, Verschure Pernette J
Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
Methods Mol Biol. 2018;1767:385-393. doi: 10.1007/978-1-4939-7774-1_21.
Single-molecule RNA fluorescent in situ hybridization (smRNA FISH) allows for the visualization, localization, and quantification of RNA transcripts within individual cells and tissues using custom-designed fluorescently labeled oligonucleotide probes. Here we describe a protocol for the preparation, imaging, and analysis of a smRNA FISH experiment that can be applied to any RNA of choice. We also provide insights as to how this powerful tool can be used to study epigenetic regulation, for example, following the epigenetic editing of genes.
单分子RNA荧光原位杂交(smRNA FISH)使用定制设计的荧光标记寡核苷酸探针,能够在单个细胞和组织中实现RNA转录本的可视化、定位及定量分析。在此,我们描述了一个smRNA FISH实验的制备、成像及分析方案,该方案可应用于任何选定的RNA。我们还提供了有关如何利用这一强大工具研究表观遗传调控的见解,例如,在基因进行表观遗传编辑之后。