RNA-Seq Analysis of Differential Splice Junction Usage and Intron Retentions by DEXSeq

PloS One
Yafang LiBin Liu

Abstract

Alternative splicing is an important biological process in the generation of multiple functional transcripts from the same genomic sequences. Differential analysis of splice junctions (SJs) and intron retentions (IRs) is helpful in the detection of alternative splicing events. In this study, we conducted differential analysis of SJs and IRs by use of DEXSeq, a Bioconductor package originally designed for differential exon usage analysis in RNA-seq data analysis. We set up an analysis pipeline including mapping of RNA-seq reads, the preparation of count tables of SJs and IRs as the input files, and the differential analysis in DEXSeq. We analyzed the public RNA-seq datasets generated from RNAi experiments on Drosophila melanogaster S2-DRSC cells to deplete RNA-binding proteins (GSE18508). The analysis confirmed previous findings on the alternative splicing of the trol and Ant2 (sesB) genes in the CG8144 (ps)-depletion experiment and identified some new alternative splicing events in other RNAi experiments. We also identified IRs that were confirmed in our SJ analysis. The proposed method used in our study can output the genomic coordinates of differentially used SJs and thus enable sequence motif search. Sequence motif search an...Continue Reading

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Citations

Jun 4, 2016·Nucleic Acids Research·Stephen W Hartley, James C Mullikin
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Datasets Mentioned

BETA
GSE18508
SRP008262

Methods Mentioned

BETA
RNA-seq

Software Mentioned

DEXSeq package
JUNCBASE
Cufflinks
IGV (
Integrated Genomic Viewer )
dexseq
SAM
SAM tools
Ensembl GRCh37
Ingenuity Pathway Analysis )

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