Blanc Valerie, Molitor Elizabeth A, Davidson Nicholas O
Gastroenterology Division, Department of Medicine, Washington University School of Medicine, Saint Louis, MO 63110, USA.
Gastroenterology Division, Department of Medicine, Washington University School of Medicine, Saint Louis, MO 63110, USA.
STAR Protoc. 2023 May 22;4(2):102313. doi: 10.1016/j.xpro.2023.102313.
RNA-binding proteins (RBPs) regulate diverse functions by interacting with target transcripts. Here we present a protocol to isolate RBP-mRNA complexes using RNA-CLIP and examine target mRNAs in association with ribosomal populations. We describe steps to identify specific RBPs and RNA targets reflecting a variety of developmental, physiological, and pathological states. This protocol enables RNP complex isolation from tissue sources (liver and small intestine) or populations of primary cells (hepatocytes), but not at a single-cell level. For complete details on the use and execution of this protocol, please refer to Blanc et al. (2014) and Blanc et al. (2021)..
RNA结合蛋白（RBPs）通过与靶转录本相互作用来调节多种功能。本文介绍了一种使用RNA交联免疫沉淀（RNA-CLIP）分离RBP-mRNA复合物并检测与核糖体群体相关的靶mRNA的方法。我们描述了识别反映各种发育、生理和病理状态的特定RBPs和RNA靶标的步骤。该方法能够从组织来源（肝脏和小肠）或原代细胞群体（肝细胞）中分离核糖核蛋白（RNP）复合物，但不能在单细胞水平上进行。有关本方法使用和实施的完整详细信息，请参考布兰克等人（2014年）和布兰克等人（2021年）的文献。
