Identifying stably expressed genes from multiple RNA-Seq data sets

We examined RNA-Seq data on 211 biological samples from 24 different Arabidopsis experiments carried out by different labs. We grouped the samples according to tissue types, and in each of the groups, we identified genes that are stably expressed across biological samples, treatment conditions, and...

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Main Authors: Bin Zhuo, Sarah Emerson, Jeff H. Chang, Yanming Di
Format: Article
Language:English
Published: PeerJ Inc. 2016-12-01
Series:PeerJ
Subjects:
Online Access:https://peerj.com/articles/2791.pdf
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author Bin Zhuo
Sarah Emerson
Jeff H. Chang
Yanming Di
author_facet Bin Zhuo
Sarah Emerson
Jeff H. Chang
Yanming Di
author_sort Bin Zhuo
collection DOAJ
description We examined RNA-Seq data on 211 biological samples from 24 different Arabidopsis experiments carried out by different labs. We grouped the samples according to tissue types, and in each of the groups, we identified genes that are stably expressed across biological samples, treatment conditions, and experiments. We fit a Poisson log-linear mixed-effect model to the read counts for each gene and decomposed the total variance into between-sample, between-treatment and between-experiment variance components. Identifying stably expressed genes is useful for count normalization and differential expression analysis. The variance component analysis that we explore here is a first step towards understanding the sources and nature of the RNA-Seq count variation. When using a numerical measure to identify stably expressed genes, the outcome depends on multiple factors: the background sample set and the reference gene set used for count normalization, the technology used for measuring gene expression, and the specific numerical stability measure used. Since differential expression (DE) is measured by relative frequencies, we argue that DE is a relative concept. We advocate using an explicit reference gene set for count normalization to improve interpretability of DE results, and recommend using a common reference gene set when analyzing multiple RNA-Seq experiments to avoid potential inconsistent conclusions.
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spelling doaj.art-3066613b2da74063bff628e09b7a46b82023-12-03T10:37:54ZengPeerJ Inc.PeerJ2167-83592016-12-014e279110.7717/peerj.2791Identifying stably expressed genes from multiple RNA-Seq data setsBin Zhuo0Sarah Emerson1Jeff H. Chang2Yanming Di3Department of Statistics, Oregon State University, Corvallis, OR, United StatesDepartment of Statistics, Oregon State University, Corvallis, OR, United StatesDepartment of Botany and Plant Pathology, Oregon State University, Corvallis, OR, United StatesDepartment of Statistics, Oregon State University, Corvallis, OR, United StatesWe examined RNA-Seq data on 211 biological samples from 24 different Arabidopsis experiments carried out by different labs. We grouped the samples according to tissue types, and in each of the groups, we identified genes that are stably expressed across biological samples, treatment conditions, and experiments. We fit a Poisson log-linear mixed-effect model to the read counts for each gene and decomposed the total variance into between-sample, between-treatment and between-experiment variance components. Identifying stably expressed genes is useful for count normalization and differential expression analysis. The variance component analysis that we explore here is a first step towards understanding the sources and nature of the RNA-Seq count variation. When using a numerical measure to identify stably expressed genes, the outcome depends on multiple factors: the background sample set and the reference gene set used for count normalization, the technology used for measuring gene expression, and the specific numerical stability measure used. Since differential expression (DE) is measured by relative frequencies, we argue that DE is a relative concept. We advocate using an explicit reference gene set for count normalization to improve interpretability of DE results, and recommend using a common reference gene set when analyzing multiple RNA-Seq experiments to avoid potential inconsistent conclusions.https://peerj.com/articles/2791.pdfStably expressed geneRNA-SeqNumerical stability measureReference gene set
spellingShingle Bin Zhuo
Sarah Emerson
Jeff H. Chang
Yanming Di
Identifying stably expressed genes from multiple RNA-Seq data sets
PeerJ
Stably expressed gene
RNA-Seq
Numerical stability measure
Reference gene set
title Identifying stably expressed genes from multiple RNA-Seq data sets
title_full Identifying stably expressed genes from multiple RNA-Seq data sets
title_fullStr Identifying stably expressed genes from multiple RNA-Seq data sets
title_full_unstemmed Identifying stably expressed genes from multiple RNA-Seq data sets
title_short Identifying stably expressed genes from multiple RNA-Seq data sets
title_sort identifying stably expressed genes from multiple rna seq data sets
topic Stably expressed gene
RNA-Seq
Numerical stability measure
Reference gene set
url https://peerj.com/articles/2791.pdf
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