Abstract

The Bru-seq Technology Platform for Nascent RNA analysis

Mats Ljungman, Professor, University of Michigan

We have developed a set of techniques that are based on the metabolic labeling of nascent RNA using bromouridine (Bru) followed by specific isolation of Bru-containing RNA and deep sequencing. In Bru-seq nascent RNA is labeled and sequenced and allows for the mapping of ongoing transcription genome-wide. In BruChase-seq, the Bru-labeled RNA is chased to obtain RNA populations of defined ages. This technique allows for the assessment of splicing kinetics as well as stability of mature RNA genome-wide. In BruUV-seq, random transcription-blocking lesions are introduced by a pulse of UV-irradiation prior to Bru labeling leading to the enhancement of signals at transcription start sites and enhancer elements. Finally, using BruDRB-seq allows for the assessment of transcription elongation rates genome-wide. The Bru-seq platform is very useful for the exploration of many aspects of transcriptional and post-transcriptional regulation genome-wide.