Protocol for high-resolution mapping of RNA-associated chromatin using chromatin isolation by RNA purification-tag

Abstract

Summary Mapping the genomic binding sites of long non-coding RNAs (lncRNAs) is essential for understanding their regulatory functions. Here, we present a protocol for integrating RNA hybridization with in situ tagmentation using chromatin isolation by RNA purification-tag (ChIRP-Tag). We describe steps for performing cell permeabilization, hybridization of biotinylated probes to target RNAs, and recruiting Protein A/G-Tn5 transposase via antibodies to catalyze in situ library construction. This protocol captures RNA-associated chromatin with high sensitivity and spatial resolution across various cell types. For complete details on the use and execution of this protocol, please refer to Fu et al.1