Université de Lorraine, CNRS, IMoPA, Nancy, France.
Methods Mol Biol. 2021;2300:89-98. doi: 10.1007/978-1-0716-1386-3_9.
Stable and transient interactions between molecules are determinant for cell function. Among those, numerous proteins contact coding and noncoding RNAs to modulate their fate and promote their activity. The identification of such interactions as well as the cellular and molecular conditions of these interactions represent key information for the characterization of the role of each partner. RNA immunoprecipitation (RIP) is the leading technique to detect in vivo the association of individual proteins with RNA species. Two main approaches exist: native RIP is largely used to identify and quantify RNA interactions, while crosslinked RIP (CLIP) may inform about direct interactions as well as their extent in the unaltered cellular condition, i.e., before cell lysis. In this chapter, both techniques applied to mammalian cells are described with a series of precautions regarding their design.
分子间的稳定和瞬时相互作用是决定细胞功能的关键。在这些相互作用中，许多蛋白质与编码和非编码 RNA 相互作用，以调节它们的命运并促进它们的活性。这些相互作用的鉴定以及这些相互作用的细胞和分子条件是表征每个伴侣作用的关键信息。RNA 免疫沉淀（RIP）是检测体内单个蛋白质与 RNA 种类结合的主要技术。主要有两种方法：天然 RIP 广泛用于鉴定和定量 RNA 相互作用，而交联 RIP（CLIP）可提供有关直接相互作用及其在未改变的细胞状态下（即在细胞裂解之前）的程度的信息。在本章中，描述了这两种技术在哺乳动物细胞中的应用，并对其设计提出了一系列注意事项。
