Use of Bru-Seq and BruChase-Seq for genome-wide assessment of the synthesis and stability of RNA.

ABSTRACT: Gene expression studies commonly examine total cellular RNA, which only provides information about its steady-state pool of RNA. It remains unclear whether differences in the steady-state reflects variable rates of transcription or RNA degradation. To specifically monitor RNA synthesis and degradation genome-wide, we developed Bru-Seq and BruChase-Seq. These assays are based on metabolic pulse-chase labeling of RNA using bromouridine (Bru). In Bru-Seq, recently labeled RNAs are sequenced to reveal spans of nascent transcription in the genome. In BruChase-Seq, cells are chased in uridine for different periods of time following Bru-labeling, allowing for the isolation of RNA populations of specific ages. Here we describe these methodologies in detail and highlight their usefulness in assessi

SUBMITTER: Paulsen MT 

PROVIDER: S-EPMC4009065 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature