BHF Centre for Cardiovascular Sciences (CVS), University of Edinburgh, Edinburgh, UK.
Department of Pathology, School for Cardiovascular Diseases (CARIM), Maastricht University, Maastricht, The Netherlands.
Methods Mol Biol. 2022;2419:659-670. doi: 10.1007/978-1-0716-1924-7_41.
In situ hybridization (ISH) is a technique for the detection of the location of RNA within a tissue of interest. This process uses oligonucleotides with complementary sequences to bind to the target RNA, and colorimetric detection to allow for the visualization of this binding. The process of ISH means that the specific location of the RNA in question can be detected, including in which cell types it is present, and the intracellular location. In the case of long noncoding RNA (lncRNA), which do not lead to the production of proteins, ISH is essential for tissue localization. Moreover, RNA abundance is often lower than for protein-coding genes, thus necessitating enhanced detection through double-digoxigenin (DIG) labeling of the probes. Here, we describe the theory and practicalities of performing ISH for lncRNA, with particular reference to vascular tissues.
原位杂交（ISH）是一种用于检测感兴趣组织中 RNA 位置的技术。该过程使用与靶 RNA 互补的寡核苷酸进行结合，并通过比色检测来允许这种结合的可视化。ISH 的过程意味着可以检测到有疑问的 RNA 的特定位置，包括它存在于哪些细胞类型中以及细胞内的位置。对于不导致蛋白质产生的长非编码 RNA（lncRNA），ISH 对于组织定位是必不可少的。此外，RNA 的丰度通常低于蛋白质编码基因，因此需要通过探针的双重地高辛（DIG）标记来增强检测。在这里，我们描述了用于 lncRNA 的 ISH 的理论和实践，特别参考了血管组织。
