Protocol to isolate RBP-mRNA complexes using RNA-CLIP and examine target mRNAs
Valerie Blanc1, Elizabeth A Molitor1, Nicholas O Davidson1
1Gastroenterology Division, Department of Medicine, Washington University School of Medicine, Saint Louis, MO 63110, USA.
Abstract:
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)1 and Blanc et al. (2021).2.


