Pseudouridine synthases modify human pre-mRNA co-transcriptionally and affect splicing

We performed pseudouridine profiling (Pseudo-seq) on 11 biological replicates of chromatin-associated RNA. We performed mRNA-seq of PUS1 knockout, PUS7 knockdown, RPUSD4 knockdown and wildtype HepG2 cells. We used an in vitro pool based strategy to assign pre-mRNA targets to individual human pseudouridine synthases (PUS) . Overall design: Pseudouridylation was measured using Pseudo-seq, a seqeuncing based technique for identifying ?s (Carlile et al. Nature 2014).