RT Journal Article
SR Electronic
T1 Transcriptome-wide interrogation of the functional intronome by spliceosome profiling
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 226894
DO 10.1101/226894
A1 Weijun Chen
A1 Jill Moore
A1 Hakan Ozadam
A1 Hennady P. Shulha
A1 Nicholas Rhind
A1 Zhiping Weng
A1 Melissa J. Moore
YR 2018
UL http://biorxiv.org/content/early/2018/03/20/226894.abstract
AB Full understanding of eukaryotic transcriptomes and how they respond to different conditions requires deep knowledge of all sites of intron excision. Although RNA-Seq provides much of this information, the low abundance of many spliced transcripts (often due to their rapid cytoplasmic decay) limits the ability of RNA-Seq alone to reveal the full repertoire of spliced species. Here we present “spliceosome profiling”, a strategy based on deep sequencing of RNAs co-purifying with late stage spliceosomes. Spliceosome profiling allows for unambiguous mapping of intron ends to single nucleotide resolution and branchpoint identification at unprecedented depths. Our data reveal hundreds of new introns in S. pombe and numerous others that were previously misannotated. By providing a means to directly interrogate sites of spliceosome assembly and catalysis genome-wide, spliceosome profiling promises to transform our understanding of RNA processing in the nucleus much like ribosome profiling has transformed our understanding mRNA translation in the cytoplasm.