Detecting protein-RNA interactions in vivo is essential for deciphering many important cellular pathways. Several methods have been described for this purpose, among which cross-linking analysis of cDNA, CRAC. This method relies on a first step of UV cross-linking of living yeast cells and several subsequent steps of purification of the protein-RNA complexes, some of which under denaturing condition. Without altering the general principle of the method, we have modified and improved the protocol, with the specific aim of sequencing the nascent RNA isolated from transcription complexes and generate high-resolution and directional transcription maps.
Keywords: CRAC (cross-linking analysis of cDNAs); Genome-wide RNAPII mapping; RNA–protein interaction in vivo; Transcription.
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.