» Articles » PMID: 30356428

Swimming Downstream: Statistical Analysis of Differential Transcript Usage Following Salmon Quantification

Overview
Journal F1000Res
Date 2018 Oct 30
PMID 30356428
Citations 74
Authors
Affiliations
Soon will be listed here.
Abstract

Detection of differential transcript usage (DTU) from RNA-seq data is an important bioinformatic analysis that complements differential gene expression analysis. Here we present a simple workflow using a set of existing R/Bioconductor packages for analysis of DTU. We show how these packages can be used downstream of RNA-seq quantification using the Salmon software package. The entire pipeline is fast, benefiting from inference steps by Salmon to quantify expression at the transcript level. The workflow includes live, runnable code chunks for analysis using DRIMSeq and DEXSeq, as well as for performing two-stage testing of DTU using the stageR package, a statistical framework to screen at the gene level and then confirm which transcripts within the significant genes show evidence of DTU. We evaluate these packages and other related packages on a simulated dataset with parameters estimated from real data.

Citing Articles

A systematic benchmark of Nanopore long-read RNA sequencing for transcript-level analysis in human cell lines.

Chen Y, Davidson N, Kei Wan Y, Yao F, Su Y, Gamaarachchi H Nat Methods. 2025; .

PMID: 40082608 DOI: 10.1038/s41592-025-02623-4.


Systematic review and meta-analysis of bulk RNAseq studies in human Alzheimer's disease brain tissue.

Heberle B, Fox K, Libermann L, Xavier S, Dallarosa G, Santos R Alzheimers Dement. 2025; 21(3):e70025.

PMID: 40042520 PMC: 11881636. DOI: 10.1002/alz.70025.


Temporal phosphoproteomics reveals circuitry of phased propagation in insulin signaling.

Turewicz M, Skagen C, Hartwig S, Majda S, Thedinga K, Herwig R Nat Commun. 2025; 16(1):1570.

PMID: 39939313 PMC: 11821911. DOI: 10.1038/s41467-025-56335-6.


A fast hybridization chain reaction method in embryos and ovaries.

Mikami K, Kozono Y, Masukawa M, Kobayashi S Fly (Austin). 2024; 19(1):2428499.

PMID: 39639000 PMC: 11633216. DOI: 10.1080/19336934.2024.2428499.


Systematic review and meta-analysis of bulk RNAseq studies in human Alzheimer's disease brain tissue.

Heberle B, Fox K, Libermann L, Xavier S, Dallarosa G, Santos R bioRxiv. 2024; .

PMID: 39574617 PMC: 11580990. DOI: 10.1101/2024.11.07.622520.


References
1.
Heller R, Manduchi E, Grant G, Ewens W . A flexible two-stage procedure for identifying gene sets that are differentially expressed. Bioinformatics. 2009; 25(8):1019-25. DOI: 10.1093/bioinformatics/btp076. View

2.
McCarthy D, Chen Y, Smyth G . Differential expression analysis of multifactor RNA-Seq experiments with respect to biological variation. Nucleic Acids Res. 2012; 40(10):4288-97. PMC: 3378882. DOI: 10.1093/nar/gks042. View

3.
Reyes A, Huber W . Alternative start and termination sites of transcription drive most transcript isoform differences across human tissues. Nucleic Acids Res. 2017; 46(2):582-592. PMC: 5778607. DOI: 10.1093/nar/gkx1165. View

4.
Van den Berge K, Soneson C, Robinson M, Clement L . stageR: a general stage-wise method for controlling the gene-level false discovery rate in differential expression and differential transcript usage. Genome Biol. 2017; 18(1):151. PMC: 5547545. DOI: 10.1186/s13059-017-1277-0. View

5.
Battle A, Brown C, Engelhardt B, Montgomery S . Genetic effects on gene expression across human tissues. Nature. 2017; 550(7675):204-213. PMC: 5776756. DOI: 10.1038/nature24277. View