miRmine dataset contains rich metadata around 304 selected publicly available,
miRNA-Seq experiments. Authors' processed the data with miRdeep2 using
annotation files from miRBase v21. Mentioned metadata is used as colData
and miRBase annotations as GRanges are used as rowRanges while preparing
this dataset as RangedSummarizedExperiment. Data used for preprocessing and
constructing the miRmine
RangedSummarizedExperiment are available in
extdata
folder. Details of this proccess could be followed in
data help file: ?miRmine
.
#library(GenomicRanges) #library(rtracklayer) #library(SummarizedExperiment) #library(Biostrings) #library(Rsamtools) ext.data <- system.file("extdata", package = "miRmine") list.files(ext.data)
Number of ranges from miRBase GFF and number of features output
by miRdeep2 are not the same (2813 vs. 2822). After closer look it turns out
that 2 rows from either tissues or cell.lines data are duplicated
(with same mature miRNA and same precursor miRNA) and 7 rows don't correspond
to mirna/precursor combination existing in miRBase v21. These rows were
removed for all samples, as seen in ?miRmine
.
To load this dataset use:
library("miRmine") data(miRmine) miRmine
You may want to further subset data on some of many colData features (Tissue, Cell Line, Disease, Sex, Instrument) or output some specifics of particular experiment(s) (Accession #, Description, Publication):
adenocarcinoma = miRmine[ , miRmine$Disease %in% c("Adenocarcinoma")] adenocarcinoma as.character(adenocarcinoma$Sample.Accession)
rowRanges data is also rich in metadata, containing all the features from miRBase hsa.gff3, with addition of actual miRNA sequence as DNAString instance. For example to read the sequence of top expressed miRNA over a subset of samples:
top.mirna = names(sort(rowSums(assays(adenocarcinoma)$counts))[1]) rowRanges(adenocarcinoma)$mirna_seq[[top.mirna]]
miRmine
could be directly used in DESeq2
(note that expression values are RPM not raw reads):
library("DESeq2") mirmine.subset = miRmine[, miRmine$Tissue %in% c("Lung", "Saliva")] mirmine.subset = SummarizedExperiment( assays = SimpleList(counts=ceiling(assays(mirmine.subset)$counts)), colData=colData(mirmine.subset), rowRanges=rowRanges(mirmine.subset), rowData=NULL ) ddsSE <- DESeqDataSet(mirmine.subset, design = ~ Tissue) ddsSE <- ddsSE[ rowSums(counts(ddsSE)) > 1, ] dds <- DESeq(ddsSE) res <- results(dds) res
sessionInfo()
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