README.md

In HuntsmanCancerInstitute/hciR: RNA-seq workflows at HCI

RNA-Seq workflows at HCI

The hciR package works best with tidyverse packages (readr, dplyr, tibble, etc.) and simplifies the code in a differential expression analysis. The package includes functions to run DESeq2 using sample and count tibbles as input, get annotated DESeq results for all pairwise comparisons and create interactive plots and other visualizations.

NOTE: Starting in hciR version 1.5, the control group should be listed first and then pairwise comparisons are selected in reverse order, so A, B, C levels will return results for C vs. B, C vs. A, and B vs. A from results_all.

Use devtools to install hciR and the hciRdata package with Ensembl annotations.

library(devtools)
install_github("HuntsmanCancerInstitute/hciR")
install_github("HuntsmanCancerInstitute/hciRdata")

The basic workflow for a mouse experiment with three groups in a trt column is listed below.

library(hciR)
samples <- read_tsv("samples.txt")
samples$trt <- factor(samples$trt, levels = c("WT", "OE", "KO"))
counts <- read_tsv("counts.txt")
counts <- filter_counts(counts, n = 5)
dds <- deseq_from_tibble(counts, samples, design = ~ trt)
rld <- r_log(dds)
plot_pca(rld, "trt", tooltip= c("id", "name"))
plot_dist(rld, "trt", na_col="white")
library(hciRdata)
res <- results_all(dds, mouse104)
plot_volcano(res[[1]])
x <- top_counts(res[[1]], rld, top=40)
plot_genes(x, "trt", scale ="row", annotation_names_col=FALSE)
write_deseq(res, dds, rld, mouse104)

Check the vignettes directory to learn more about the package. The Pasilla vignette runs through an analysis with a single contrast and Liver includes an interaction model and gene set enrichment. The Ensembl file has details on loading annotations.

The hciRscripts package wraps functions like read_featureCounts to run on the command line. See the hciR scripts file for more details.

HuntsmanCancerInstitute/hciR documentation built on March 26, 2024, 3:09 a.m.