Home LiteratureArticle Details
PMID: 23821648 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Data-based filtering for replicated high-throughput transcriptome sequencing experiments.

Bioinformatics (Oxford, England) ·Vol. 29 ·No. 17 ·2013-09-01 ·Pages 2146-52

Rau A, Gallopin M, Celeux G, Jaffrézic F

Abstract

RNA sequencing is now widely performed to study differential expression among experimental conditions. As tests are performed on a large number of genes, stringent false-discovery rate control is required at the expense of detection power. Ad hoc filtering techniques are regularly used to moderate this correction by removing genes with low signal, with little attention paid to their impact on downstream analyses. We propose a data-driven method based on the Jaccard similarity index to calculate a filtering threshold for replicated RNA sequencing data. In comparisons with alternative data filters regularly used in practice, we demonstrate the effectiveness of our proposed method to correctly filter lowly expressed genes, leading to increased detection power for moderately to highly expressed genes. Interestingly, this data-driven threshold varies among experiments, highlighting the interest of the method proposed here. The proposed filtering method is implemented in the R package HTSFilter available on Bioconductor.

MeSH Terms
Animals Gene Expression Profiling/methods High-Throughput Nucleotide Sequencing/methods Humans Mice Sequence Analysis, RNA/methods
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Rau Andrea
INRA, UMR1313 Génétique animale et biologie intégrative, 78352 Jouy-en-Josas, France. [email protected]
Gallopin Mélina
Celeux Gilles
Jaffrézic Florence
References (22)
22 references, click to expand
  1. An overview of Ensembl.
    Genome Res. 2004 May;14(5):925-8 PMID: 15078858
  2. Independent filtering increases detection power for high-throughput experiments.
    Proc Natl Acad Sci U S A. 2010 May 25;107(21):9546-51 PMID: 20460310
  3. SNP discovery in the bovine milk transcriptome using RNA-Seq technology.
    Mamm Genome. 2010 Dec;21(11-12):592-8 PMID: 21057797
  4. From RNA-seq reads to differential expression results.
    Genome Biol. 2010;11(12):220 PMID: 21176179
  5. Characterization and improvement of RNA-Seq precision in quantitative transcript expression profiling.
    Bioinformatics. 2011 Jul 1;27(13):i383-91 PMID: 21685096
  6. A scaling normalization method for differential expression analysis of RNA-seq data.
    Genome Biol. 2010;11(3):R25 PMID: 20196867
  7. Differential expression analysis for sequence count data.
    Genome Biol. 2010;11(10):R106 PMID: 20979621
  8. DEGseq: an R package for identifying differentially expressed genes from RNA-seq data.
    Bioinformatics. 2010 Jan 1;26(1):136-8 PMID: 19855105
  9. An abundance of ubiquitously expressed genes revealed by tissue transcriptome sequence data.
    PLoS Comput Biol. 2009 Dec;5(12):e1000598 PMID: 20011106
  10. Removing technical variability in RNA-seq data using conditional quantile normalization.
    Biostatistics. 2012 Apr;13(2):204-16 PMID: 22285995
  11. GC-content normalization for RNA-Seq data.
    BMC Bioinformatics. 2011 Dec 17;12:480 PMID: 22177264
  12. Essential role of microphthalmia transcription factor for DNA replication, mitosis and genomic stability in melanoma.
    Oncogene. 2011 May 19;30(20):2319-32 PMID: 21258399
  13. A global view of gene activity and alternative splicing by deep sequencing of the human transcriptome.
    Science. 2008 Aug 15;321(5891):956-60 PMID: 18599741
  14. Transcript length bias in RNA-seq data confounds systems biology.
    Biol Direct. 2009 Apr 16;4:14 PMID: 19371405
  15. ReCount: a multi-experiment resource of analysis-ready RNA-seq gene count datasets.
    BMC Bioinformatics. 2011 Nov 16;12:449 PMID: 22087737
  16. A comparison of single molecule and amplification based sequencing of cancer transcriptomes.
    PLoS One. 2011 Mar 01;6(3):e17305 PMID: 21390249
  17. Bioconductor: open software development for computational biology and bioinformatics.
    Genome Biol. 2004;5(10):R80 PMID: 15461798
  18. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data.
    Bioinformatics. 2010 Jan 1;26(1):139-40 PMID: 19910308
  19. Mapping and quantifying mammalian transcriptomes by RNA-Seq.
    Nat Methods. 2008 Jul;5(7):621-8 PMID: 18516045
  20. EnsMart: a generic system for fast and flexible access to biological data.
    Genome Res. 2004 Jan;14(1):160-9 PMID: 14707178
  21. A comprehensive evaluation of normalization methods for Illumina high-throughput RNA sequencing data analysis.
    Brief Bioinform. 2013 Nov;14(6):671-83 PMID: 22988256
  22. Evaluating gene expression in C57BL/6J and DBA/2J mouse striatum using RNA-Seq and microarrays.
    PLoS One. 2011 Mar 24;6(3):e17820 PMID: 21455293
Article Info
Journal
Bioinformatics (Oxford, England)
Abbr.
Bioinformatics
ISSN
1367-4811
Published
2013-09-01
Epub
2013-00-02
Pages
2146-52
Language
English
Region
England
NLM ID
9808944
PMCID
PMC3740625
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]