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Fast and accurate differential transcript usage by testing equivalence class counts

View ORCID ProfileMarek Cmero, View ORCID ProfileNadia M Davidson, View ORCID ProfileAlicia Oshlack
doi: https://doi.org/10.1101/501106
Marek Cmero
1Murdoch Children’s Research Institute, Flemington Road, Parkville, Australia
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Nadia M Davidson
1Murdoch Children’s Research Institute, Flemington Road, Parkville, Australia
2School of Biosciences, Faculty of Science, University of Melbourne, Melbourne, Victoria, Australia
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Alicia Oshlack
1Murdoch Children’s Research Institute, Flemington Road, Parkville, Australia
2School of Biosciences, Faculty of Science, University of Melbourne, Melbourne, Victoria, Australia
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  • For correspondence: alicia.oshlack@mcri.edu.au
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Abstract

RNA sequencing has enabled high-throughput and fine-grained quantitative analyses of the transcriptome. While differential gene expression is the most widely used application of this technology, RNA-seq data also has the resolution to infer differential transcript usage (DTU), which can elucidate the role of different transcript isoforms between experimental conditions, cell types or tissues. DTU has typically been inferred from exon-count data, which has issues with assigning reads unambiguously to counting bins, and requires alignment of reads to the genome. Recently, approaches have emerged that use transcript quantifications estimates directly for DTU. Transcript counts can be inferred from ‘pseudo’ or lightweight aligners, which are significantly faster than traditional genome alignment. However, recent evaluations show lower sensitivity in DTU analysis. Transcript abundances are estimated from equivalence classes (ECs), which determine the transcripts that any given read is compatible with. Here we propose performing DTU testing directly on equivalence class read counts. We evaluate this approach on simulated human and drosophila data, as well as on a real dataset through subset testing. We find that ECs counts have similar sensitivity and false discovery rates as exon-level counts but can be generated in a fraction of the time through the use of pseudo-aligners. We posit that equivalent class counts is a natural unit on which to perform many types of analysis.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted December 19, 2018.
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Fast and accurate differential transcript usage by testing equivalence class counts
Marek Cmero, Nadia M Davidson, Alicia Oshlack
bioRxiv 501106; doi: https://doi.org/10.1101/501106
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Fast and accurate differential transcript usage by testing equivalence class counts
Marek Cmero, Nadia M Davidson, Alicia Oshlack
bioRxiv 501106; doi: https://doi.org/10.1101/501106

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