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| Original file line number | Diff line number | Diff line change |
|---|---|---|
| @@ -1,94 +1,143 @@ | ||
| #!/usr/bin/env Rscript | ||
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| # Written by Lorena Pantano and released under the MIT license. | ||
| # Script for importing and processing transcript-level quantifications. | ||
| # Written by Lorena Pantano, later modified by Jonathan Manning, and released | ||
| # under the MIT license. | ||
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| # Loading required libraries | ||
| library(SummarizedExperiment) | ||
| library(tximport) | ||
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| args = commandArgs(trailingOnly=TRUE) | ||
| # Parsing command line arguments | ||
| args <- commandArgs(trailingOnly=TRUE) | ||
| if (length(args) < 4) { | ||
| stop("Usage: tximport.r <coldata> <path> <sample_name> <quant_type> <tx2gene_path>", call.=FALSE) | ||
| stop("Usage: tximport.r <coldata_path> <path> <prefix> <quant_type> <tx2gene_path>", | ||
| call.=FALSE) | ||
| } | ||
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| coldata = args[1] | ||
| path = args[2] | ||
| sample_name = args[3] | ||
| quant_type = args[4] | ||
| tx2gene_path = args[5] | ||
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| prefix = sample_name | ||
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| info = file.info(tx2gene_path) | ||
| if (info$size == 0) { | ||
| tx2gene = NULL | ||
| } else { | ||
| rowdata = read.csv(tx2gene_path, sep="\t", header = FALSE) | ||
| colnames(rowdata) = c("tx", "gene_id", "gene_name") | ||
| tx2gene = rowdata[,1:2] | ||
| # Assigning command line arguments to variables | ||
| coldata_path <- args[1] | ||
| path <- args[2] | ||
| prefix <- args[3] | ||
| quant_type <- args[4] | ||
| tx2gene_path <- args[5] | ||
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| ## Functions | ||
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| # Build a table from a SummarizedExperiment object | ||
| build_table <- function(se.obj, slot) { | ||
| cbind(rowData(se.obj)[,1:2], assays(se.obj)[[slot]]) | ||
| } | ||
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| pattern <- ifelse(quant_type == "kallisto", "abundance.tsv", "quant.sf") | ||
| fns = list.files(path, pattern = pattern, recursive = T, full.names = T) | ||
| names = basename(dirname(fns)) | ||
| names(fns) = names | ||
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| if (file.exists(coldata)) { | ||
| coldata = read.csv(coldata, sep="\t") | ||
| coldata = coldata[match(names, coldata[,1]),] | ||
| coldata = cbind(files = fns, coldata) | ||
| } else { | ||
| message("ColData not available: ", coldata) | ||
| coldata = data.frame(files = fns, names = names) | ||
| # Write a table to a file with given parameters | ||
| write_se_table <- function(params) { | ||
| file_name <- paste0(prefix, ".", params$suffix) | ||
| write.table(build_table(params$obj, params$slot), file_name, | ||
| sep="\t", quote=FALSE, row.names = FALSE) | ||
| } | ||
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| dropInfReps = quant_type == "kallisto" | ||
| # Read transcript metadata from a given path | ||
| read_transcript_info <- function(tinfo_path){ | ||
| info <- file.info(tinfo_path) | ||
| if (info$size == 0) { | ||
| stop("tx2gene file is empty") | ||
| } | ||
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| transcript_info <- read.csv(tinfo_path, sep="\t", header = FALSE, | ||
| col.names = c("tx", "gene_id", "gene_name")) | ||
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| txi = tximport(fns, type = quant_type, txOut = TRUE, dropInfReps = dropInfReps) | ||
| rownames(coldata) = coldata[["names"]] | ||
| extra = setdiff(rownames(txi[[1]]), as.character(rowdata[["tx"]])) | ||
| if (length(extra) > 0) { | ||
| rowdata = rbind(rowdata, data.frame(tx=extra, gene_id=extra, gene_name=extra)) | ||
| extra <- setdiff(rownames(txi[[1]]), as.character(transcript_info[["tx"]])) | ||
| transcript_info <- rbind(transcript_info, data.frame(tx=extra, gene_id=extra, gene_name=extra)) | ||
| transcript_info <- transcript_info[match(rownames(txi[[1]]), transcript_info[["tx"]]), ] | ||
| rownames(transcript_info) <- transcript_info[["tx"]] | ||
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| list(transcript = transcript_info, | ||
| gene = unique(transcript_info[,2:3]), | ||
| tx2gene = transcript_info[,1:2]) | ||
| } | ||
| rowdata = rowdata[match(rownames(txi[[1]]), as.character(rowdata[["tx"]])),] | ||
| rownames(rowdata) = rowdata[["tx"]] | ||
| se = SummarizedExperiment(assays = list(counts = txi[["counts"]], abundance = txi[["abundance"]], length = txi[["length"]]), | ||
| colData = DataFrame(coldata), | ||
| rowData = rowdata) | ||
| if (!is.null(tx2gene)) { | ||
| gi = summarizeToGene(txi, tx2gene = tx2gene) | ||
| gi.ls = summarizeToGene(txi, tx2gene = tx2gene, countsFromAbundance = "lengthScaledTPM") | ||
| gi.s = summarizeToGene(txi, tx2gene = tx2gene, countsFromAbundance = "scaledTPM") | ||
| growdata = unique(rowdata[,2:3]) | ||
| growdata = growdata[match(rownames(gi[[1]]), growdata[["gene_id"]]),] | ||
| rownames(growdata) = growdata[["tx"]] | ||
| gse = SummarizedExperiment(assays = list(counts = gi[["counts"]], abundance = gi[["abundance"]], length = gi[["length"]]), | ||
| colData = DataFrame(coldata), | ||
| rowData = growdata) | ||
| gse.ls = SummarizedExperiment(assays = list(counts = gi.ls[["counts"]], abundance = gi.ls[["abundance"]], length = gi.ls[["length"]]), | ||
| colData = DataFrame(coldata), | ||
| rowData = growdata) | ||
| gse.s = SummarizedExperiment(assays = list(counts = gi.s[["counts"]], abundance = gi.s[["abundance"]], length = gi.s[["length"]]), | ||
| colData = DataFrame(coldata), | ||
| rowData = growdata) | ||
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| # Read and process sample/column data from a given path | ||
| read_coldata <- function(coldata_path){ | ||
| if (file.exists(coldata_path)) { | ||
| coldata <- read.csv(coldata_path, sep="\t") | ||
| coldata <- coldata[match(names, coldata[,1]),] | ||
| coldata <- cbind(files = fns, coldata) | ||
| } else { | ||
| message("ColData not available: ", coldata_path) | ||
| coldata <- data.frame(files = fns, names = names) | ||
| } | ||
| rownames(coldata) <- coldata[["names"]] | ||
| } | ||
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| build_table = function(se.obj, slot) { | ||
| cbind(rowData(se.obj)[,1:2], assays(se.obj)[[slot]]) | ||
| # Create a SummarizedExperiment object with given data | ||
| create_summarized_experiment <- function(counts, abundance, length, col_data, row_data) { | ||
| SummarizedExperiment(assays = list(counts = counts, abundance = abundance, length = length), | ||
| colData = col_data, | ||
| rowData = row_data) | ||
| } | ||
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| if(exists("gse")){ | ||
| write.table(build_table(gse, "abundance"), paste(c(prefix, "gene_tpm.tsv"), collapse="."), sep="\t", quote=FALSE, row.names = FALSE) | ||
| write.table(build_table(gse, "counts"), paste(c(prefix, "gene_counts.tsv"), collapse="."), sep="\t", quote=FALSE, row.names = FALSE) | ||
| write.table(build_table(gse.ls, "abundance"), paste(c(prefix, "gene_tpm_length_scaled.tsv"), collapse="."), sep="\t", quote=FALSE, row.names = FALSE) | ||
| write.table(build_table(gse.ls, "counts"), paste(c(prefix, "gene_counts_length_scaled.tsv"), collapse="."), sep="\t", quote=FALSE, row.names = FALSE) | ||
| write.table(build_table(gse.s, "abundance"), paste(c(prefix, "gene_tpm_scaled.tsv"), collapse="."), sep="\t", quote=FALSE, row.names = FALSE) | ||
| write.table(build_table(gse.s, "counts"), paste(c(prefix, "gene_counts_scaled.tsv"), collapse="."), sep="\t", quote=FALSE, row.names = FALSE) | ||
| # Main script starts here | ||
|
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| # Define pattern for file names based on quantification type | ||
| pattern <- ifelse(quant_type == "kallisto", "abundance.tsv", "quant.sf") | ||
| fns <- list.files(path, pattern = pattern, recursive = T, full.names = T) | ||
| names <- basename(dirname(fns)) | ||
| names(fns) <- names | ||
| dropInfReps <- quant_type == "kallisto" | ||
|
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| # Import transcript-level quantifications | ||
| txi <- tximport(fns, type = quant_type, txOut = TRUE, dropInfReps = dropInfReps) | ||
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| # Read transcript and sample data | ||
| transcript_info <- read_transcript_info(tx2gene_path) | ||
| coldata <- read_coldata(coldata_path) | ||
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| # Create initial SummarizedExperiment object | ||
| se <- create_summarized_experiment(txi[["counts"]], txi[["abundance"]], txi[["length"]], | ||
| DataFrame(coldata), transcript_info$transcript) | ||
|
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| # Setting parameters for writing tables | ||
| params <- list( | ||
| list(obj = se, slot = "abundance", suffix = "transcript_tpm.tsv"), | ||
| list(obj = se, slot = "counts", suffix = "transcript_counts.tsv"), | ||
| list(obj = se, slot = "length", suffix = "transcript_lengths.tsv") | ||
| ) | ||
|
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| # Process gene-level data if tx2gene mapping is available | ||
| if ("tx2gene" %in% names(transcript_info) && !is.null(transcript_info$tx2gene)) { | ||
| tx2gene <- transcript_info$tx2gene | ||
| gi <- summarizeToGene(txi, tx2gene = tx2gene) | ||
| gi.ls <- summarizeToGene(txi, tx2gene = tx2gene, countsFromAbundance = "lengthScaledTPM") | ||
| gi.s <- summarizeToGene(txi, tx2gene = tx2gene, countsFromAbundance = "scaledTPM") | ||
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| gene_info <- transcript_info$gene[match(rownames(gi[[1]]), transcript_info$gene[["gene_id"]]),] | ||
| rownames(gene_info) <- gene_info[["tx"]] | ||
|
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| col_data_frame <- DataFrame(coldata) | ||
|
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| # Create gene-level SummarizedExperiment objects | ||
| gse <- create_summarized_experiment(gi[["counts"]], gi[["abundance"]], gi[["length"]], | ||
| col_data_frame, gene_info) | ||
| gse.ls <- create_summarized_experiment(gi.ls[["counts"]], gi.ls[["abundance"]], gi.ls[["length"]], | ||
| col_data_frame, gene_info) | ||
| gse.s <- create_summarized_experiment(gi.s[["counts"]], gi.s[["abundance"]], gi.s[["length"]], | ||
| col_data_frame, gene_info) | ||
|
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| params <- c(params, list( | ||
| list(obj = gse, slot = "length", suffix = "gene_lengths.tsv"), | ||
| list(obj = gse, slot = "abundance", suffix = "gene_tpm.tsv"), | ||
| list(obj = gse, slot = "counts", suffix = "gene_counts.tsv"), | ||
| list(obj = gse.ls, slot = "abundance", suffix = "gene_tpm_length_scaled.tsv"), | ||
| list(obj = gse.ls, slot = "counts", suffix = "gene_counts_length_scaled.tsv"), | ||
| list(obj = gse.s, slot = "abundance", suffix = "gene_tpm_scaled.tsv"), | ||
| list(obj = gse.s, slot = "counts", suffix = "gene_counts_scaled.tsv") | ||
| )) | ||
| } | ||
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| write.table(build_table(se, "abundance"), paste(c(prefix, "transcript_tpm.tsv"), collapse="."), sep="\t", quote=FALSE, row.names = FALSE) | ||
| write.table(build_table(se, "counts"), paste(c(prefix, "transcript_counts.tsv"), collapse="."), sep="\t", quote=FALSE, row.names = FALSE) | ||
| # Writing tables for each set of parameters | ||
| done <- lapply(params, write_se_table) | ||
|
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| # Print sessioninfo to standard out | ||
| # Output session information and citations | ||
| citation("tximeta") | ||
| sessionInfo() | ||
|
|
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