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Nat Biotechnol. 2014 Apr;32(4):381-6. doi: 10.1038/nbt.2859. Epub 2014 Mar 23.

The dynamics and regulators of cell fate decisions are revealed by pseudotemporal ordering of single cells.

Author information

1
1] Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA. [2] The Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA. [3].
2
1] Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA. [2] The Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA. [3] Harvard Stem Cell Institute, Harvard University, Cambridge, Massachusetts, USA. [4].
3
The Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
4
Fluidigm Corporation, South San Francisco, California, USA.
5
1] Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA. [2] The Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
6
1] Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA. [2] The Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA. [3] Harvard Stem Cell Institute, Harvard University, Cambridge, Massachusetts, USA.
7
1] Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA. [2] The Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA. [3] Department of Pathology, Beth Israel Deaconess Medical Center, Boston, Massachusetts, USA.

Abstract

Defining the transcriptional dynamics of a temporal process such as cell differentiation is challenging owing to the high variability in gene expression between individual cells. Time-series gene expression analyses of bulk cells have difficulty distinguishing early and late phases of a transcriptional cascade or identifying rare subpopulations of cells, and single-cell proteomic methods rely on a priori knowledge of key distinguishing markers. Here we describe Monocle, an unsupervised algorithm that increases the temporal resolution of transcriptome dynamics using single-cell RNA-Seq data collected at multiple time points. Applied to the differentiation of primary human myoblasts, Monocle revealed switch-like changes in expression of key regulatory factors, sequential waves of gene regulation, and expression of regulators that were not known to act in differentiation. We validated some of these predicted regulators in a loss-of function screen. Monocle can in principle be used to recover single-cell gene expression kinetics from a wide array of cellular processes, including differentiation, proliferation and oncogenic transformation.

PMID:
24658644
PMCID:
PMC4122333
DOI:
10.1038/nbt.2859
[Indexed for MEDLINE]
Free PMC Article
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