Abstract

The phenotypic diversity of cells is governed by a complex equilibrium between their genetic identity and their environmental interactions: Understanding the dynamics of gene expression is a fundamental question of biology. However, analysing time-course transcriptomic data raises unique challenging statistical and computational questions, requiring the development of novel methods and software. This workflow provides a step-by-step tutorial of the methodology used to analyse time-course data: (1) quality control and normalization of the dataset; (2) differential expression analysis using functional data analysis; (3) clustering of time-course data; (4) interpreting clusters with GO term and KEGG pathway enrichment analysis. As a case study, we apply this workflow to time-course transcriptomic data from mice exposed to four strains of influenza to showcase every step of the pipeline.

Highlights

  • Gene expression studies provide simultaneous quantification of the level of mRNA from all genes in a sample

  • A new variety of time course studies have come from single-cell sequencing experiments (Habib et al, 2016; Shalek et al, 2014; Trapnell et al, 2014) which can sequence single cells at different stages of development; in this case, the time point is the stage of the cell in the process of development -- a value that is not know but estimated from the data as its “pseudo-time.”

  • We find in practice that the global analysis simplifies analysis and interpretation of longer time courses data, with per-time point analysis reserved for interesting comparisons of individual time-points

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Summary

11 Dec 2020 report

1. Michael I. Love , University of North Carolina at Chapel Hill, Chapel Hill, USA Any reports and responses or comments on the article can be found at the end of the article. Keywords time-course gene expression data, clustering, differential expression, workflow This article is included in the Bioconductor gateway. This article is included in the RPackage gateway.

Introduction
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19 GO:0090596 sensory organ morphogenesis
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