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dc.contributor.authorChen, Y
dc.contributor.authorPal, B
dc.contributor.authorVisvader, JE
dc.contributor.authorSmyth, GK
dc.date.accessioned2020-12-18T03:51:56Z
dc.date.available2020-12-18T03:51:56Z
dc.date.issued2017
dc.identifier.citationChen, Y., Pal, B., Visvader, J. E. & Smyth, G. K. (2017). Differential methylation analysis of reduced representation bisulfite sequencing experiments using edgeR.. F1000Res, 6, pp.2055-. https://doi.org/10.12688/f1000research.13196.2.
dc.identifier.issn2046-1402
dc.identifier.urihttp://hdl.handle.net/11343/255952
dc.description.abstractCytosine methylation is an important DNA epigenetic modification. In vertebrates, methylation occurs at CpG sites, which are dinucleotides where a cytosine is immediately followed by a guanine in the DNA sequence from 5' to 3'. When located in the promoter region of a gene, DNA methylation is often associated with transcriptional silencing of the gene. Aberrant DNA methylation is associated with the development of various diseases such as cancer. Bisulfite sequencing (BS-seq) is the current "gold-standard" technology for high-resolution profiling of DNA methylation. Reduced representation bisulfite sequencing (RRBS) is an efficient form of BS-seq that targets CpG-rich DNA regions in order to save sequencing costs. A typical bioinformatics aim is to identify CpGs that are differentially methylated (DM) between experimental conditions. This workflow demonstrates that differential methylation analysis of RRBS data can be conducted using software and methodology originally developed for RNA-seq data. The RNA-seq pipeline is adapted to methylation by adding extra columns to the design matrix to account for read coverage at each CpG, after which the RRBS and RNA-seq pipelines are almost identical. This approach is statistically natural and gives analysts access to a rich collection of analysis tools including generalized linear models, gene set testing and pathway analysis. The article presents a complete start to finish case study analysis of RRBS profiles of different cell populations from the mouse mammary gland using the Bioconductor package edgeR. We show that lineage-committed cells are typically hyper-methylated compared to progenitor cells and this is true on all the autosomes but not the sex chromosomes. We demonstrate a strong negative correlation between methylation of promoter regions and gene expression as measured by RNA-seq for the same cell types, showing that methylation is a regulatory mechanism involved in epithelial linear commitment.
dc.languageeng
dc.publisherF1000 Research Ltd
dc.rights.urihttps://creativecommons.org/licenses/by/4.0
dc.titleDifferential methylation analysis of reduced representation bisulfite sequencing experiments using edgeR.
dc.typeJournal Article
dc.identifier.doi10.12688/f1000research.13196.2
melbourne.affiliation.departmentSchool of Mathematics and Statistics
melbourne.affiliation.departmentMedical Biology (W.E.H.I.)
melbourne.source.titleF1000Research
melbourne.source.volume6
melbourne.source.pages2055-
dc.rights.licenseCC BY
melbourne.elementsid1290553
melbourne.openaccess.pmchttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC5747346
melbourne.contributor.authorPal, Bhupinder
melbourne.contributor.authorSmyth, Gordon
melbourne.contributor.authorVisvader, Jane
dc.identifier.eissn2046-1402
melbourne.accessrightsOpen Access


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