Capture of RNA-binding proteins across mouse tissues using HARD-AP

Capture of RNA-binding proteins across mouse tissues using HARD-AP

Blog Article

Abstract RNA-binding proteins (RBPs) modulate all aspects of RNA metabolism, but a comprehensive picture of RBP expression across tissues is lacking.Here, we describe our development of the method we call HARD-AP that robustly retrieves RBPs and tightly associated RNA regulatory complexes from cultured esp sd-2 cells and fresh tissues.We successfully use HARD-AP to establish a comprehensive atlas of RBPs across mouse primary organs.We then systematically map RNA-binding sites of these RBPs using machine learning-based modeling.Notably, the hp 15-da0008ca modeling reveals that the LIM domain as an RNA-binding domain in many RBPs.

We validate the LIM-domain-only protein Csrp1 as a tissue-dependent RNA binding protein.Taken together, HARD-AP is a powerful approach that can be used to identify RBPomes from any type of sample, allowing comprehensive and physiologically relevant networks of RNA-protein interactions.