Transcriptome-wide analysis of the functional intronome using spliceosome profiling

摘要: Full understanding of eukaryotic transcriptomes and how they respond to different conditions requires deep knowledge all sites intron excision. Although RNA-Seq provides much this information, the low abundance many spliced transcripts (often due their rapid cytoplasmic decay) limits ability alone reveal full repertoire species. Here we present "spliceosome profiling", a strategy based on sequencing RNAs copurifying with late stage spliceosomes. Spliceosome profiling allows for unambiguous mapping ends single nucleotide resolution branchpoint identification at unprecedented depths. Our data hundreds new introns in S. pombe numerous others that were previously misannotated. By providing means directly interrogate spliceosome assembly catalysis genome-wide, may well transform our RNA processing nucleus as ribosome is transforming mRNA translation cytoplasm.