Throughout its life cycle, each RNA will interact dynamically with numerous RNA-binding proteins (RBPs) that determine how the RNA is processed, where it is localized, whether it is translated and finally how it is degraded. In turn, RBPs can interact with hundreds to thousands of cellular RNAs by recognizing certain sequence motifs or structural elements. Understanding these interactions in a genome-wide manner is crucial for gaining a complete picture of an RBP’s specificity and function.
Individual-nucleotide resolution UV crosslinking and immunoprecipitation (iCLIP) has established itself as a leading method to map the RNA interaction sites of a single RBP-of-interest with high resolution and in a quantitative manner. The major objective of this EMBO Practical Course is to disseminate the knowledge and proficiency in this technically challenging ribonomics method. We will guide the participants through the complete protocol, placing special emphasis on critical optimisation steps and controls. We will further discuss the bioinformatics strategies for analysis of iCLIP data. The hands-on sessions in the laboratory will be complemented by presentations by established scientists that will share their experience on using iCLIP to answer different biological questions.