使用 RNA 接近连接分析评估原位 RNA-RNA 相互作用的方案。
Protocol to assess RNA-RNA interactions in situ using an RNA-proximity ligation assay.
机构信息
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Institute for Immunology, University of Pennsylvania, Philadelphia, PA 19104, USA.
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Institute for Immunology, University of Pennsylvania, Philadelphia, PA 19104, USA.
出版信息
STAR Protoc. 2022 Dec 16;3(4):101892. doi: 10.1016/j.xpro.2022.101892. Epub 2022 Dec 1.
Here, we describe a protocol to assess RNA-RNA interactions in situ using an adapted proximity ligation assay (PLA). We detail steps to perform RNA-probe hybridization, in situ rolling circle amplification, and immunofluorescence confocal microscopy. With these tools, it is possible to detect and characterize the intracellular localization of interacting RNA pairs using small cell numbers. This protocol provides a targeted approach to understanding RNA-RNA interactions in intact cells that can complement other established deep-sequencing-based approaches. For complete details on the use and execution of this protocol, please refer to Basavappa et al. (2022)..
在这里，我们描述了一种使用改良的邻近连接分析 (PLA) 来评估 RNA-RNA 相互作用的方案。我们详细介绍了进行 RNA 探针杂交、原位滚环扩增和免疫荧光共焦显微镜的步骤。有了这些工具，就可以使用少量细胞来检测和描述相互作用的 RNA 对的细胞内定位。该方案为深入了解完整细胞中的 RNA-RNA 相互作用提供了一种靶向方法，可以补充其他已建立的基于深度测序的方法。如需了解本方案的使用和执行的完整详细信息，请参考 Basavappa 等人 (2022)。
Zotero 插件