Characterization of the single-cell transcriptional landscape by highly multiplex RNA-seq

  1. Sten Linnarsson1,5
  1. 1Laboratory for Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, SE-171 77 Stockholm, Sweden;
  2. 2Department of Neuroscience, Karolinska Institutet, SE-171 77 Stockholm, Sweden;
  3. 3Illumina Inc., San Diego, California 92121, USA
    1. 4 These authors contributed equally to this work.

    Abstract

    Our understanding of the development and maintenance of tissues has been greatly aided by large-scale gene expression analysis. However, tissues are invariably complex, and expression analysis of a tissue confounds the true expression patterns of its constituent cell types. Here we describe a novel strategy to access such complex samples. Single-cell RNA-seq expression profiles were generated, and clustered to form a two-dimensional cell map onto which expression data were projected. The resulting cell map integrates three levels of organization: the whole population of cells, the functionally distinct subpopulations it contains, and the single cells themselves—all without need for known markers to classify cell types. The feasibility of the strategy was demonstrated by analyzing the transcriptomes of 85 single cells of two distinct types. We believe this strategy will enable the unbiased discovery and analysis of naturally occurring cell types during development, adult physiology, and disease.

    Footnotes

    • Received May 27, 2010.
    • Accepted April 19, 2011.
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