Cited by 1,170Single-cell and spatial transcriptomicsGenomics and Phylogenetic StudiesGene expression and cancer classification

Computational and analytical challenges in single-cell transcriptomics

Stegle, Oliver, Teichmann, Sarah A., Marioni, John C.·2015·Nature Reviews Genetics

Abstract

The development of high-throughput RNA sequencing (RNA-seq) at the single-cell level has already led to profound new discoveries in biology, ranging from the identification of novel cell types to the study of global patterns of stochastic gene expression. Alongside the technological breakthroughs that have facilitated the large-scale generation of single-cell transcriptomic data, it is important to consider the specific computational and analytical challenges that still have to be overcome. Although some tools for analysing RNA-seq data from bulk cell populations can be readily applied to single-cell RNA-seq data, many new computational strategies are required to fully exploit this data type and to enable a comprehensive yet detailed study of gene expression at the single-cell level.

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Crossref · 10.1038/nrg3833

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89 of 89 references verified0 citation errors · 0 not checkable

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  • Single-cell RNA-seq data exhibits significant cell-to-cell variation due to technical factors, including the number of molecules detected in each cell, which can confound biological heterogeneity with technical effects.

    “cell-to-cell variation in technical factors can also confound these results [1, 2].”
  • Multiple replicates of C1 collections from the same biological condition are necessary to facilitate estimation of technical variability even with ERCC spike-in controls.

    “Consequently, multiple replicates of C1 collections from the same biological condition are necessary to facilitate estimation of technical variability even with the presence of ERCC spike-in controls10.”
  • ERCCs can be problematic for modeling endogenous mRNAs, potentially due to their shorter length, shorter poly-A tail, and missing 5' cap.

    “It has been noted before that ERCCs can be problematic for modeling endogenous mRNAs (Risso et al., 2014), potentially due to their shorter length, shorter poly-A tail, and their missing 5 0 cap (Gr € un and van Oudenaarden, 2015; Stegle et al., 2015).”

    From Comparative Analysis of Single-Cell RNA Sequencing Methods (2017) · cites this paper as Stegle et al., 2015

  • Extra Poisson variability includes biological variation and technical variation, and technical variation includes noise introduced by amplification.

    “This extra Poisson variability includes biological variation (assumed to be the same across methods in our data) and technical variation, and the latter includes noise introduced by amplification (Brennecke et al., 2013; Gr € un et al., 2014; Stegle et al., 2015).”

    From Comparative Analysis of Single-Cell RNA Sequencing Methods (2017) · cites this paper as Stegle et al., 2015

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Authors
Stegle, O., Teichmann, S. A., & Marioni, J. C.
Affiliations
European Bioinformatics Institute (GB)
Corresponding author
Marioni, John C.
Venue
Nature Reviews Genetics
Published
2015-01-28
Volume
16
Issue
3
Pages
133-145
ISSN
  • 1471-0056 (ISSN-L)
  • 1471-0064
OpenAlex
W1988391018
Cited by
1,170
References
90
Citations by year
2026: 40 · 2025: 83 · 2024: 85 · 2023: 100 · 2022: 106 · 2021: 122 · 2020: 130 · 2019: 132 · 2018: 112 · 2017: 120 · 2016: 102 · 2015: 35 · 2014: 1
FWCI
11.92
Type
journal article
Language
en
Open access
closed
Locations
  • Nature Reviews Genetics (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Phylogenetic Studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene expression and cancer classification
Keywords
single-cell RNA sequencing, single-cell transcriptomics, computational challenges, data analysis, stochastic gene expression, cell type annotation, gene expression analysis
MeSH terms
Animals, Cell Differentiation, Eukaryotic Cells, Humans, Quality Control, RNA, Messenger, Software, Gene Expression Profiling, Gene Regulatory Networks, Single-Cell Analysis, High-Throughput Nucleotide Sequencing, Transcriptome

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