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Transcription factor profiling in individual hematopoietic progenitors by digital RT-PCR

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Author

  • Luigi Warren
  • David Bryder
  • Irving L. Weissman
  • Stephen R. Quake

Summary, in English

We report here a systematic, quantitative population analysis of transcription factor expression within developmental progenitors, made possible by a microfluidic chip-based "digital RT-PCR" assay that can count template molecules in cDNA samples prepared from single cells. In a survey encompassing five classes of early hematopoietic precursor, we found markedly heterogeneous expression of the transcription factor PU.1 in hematopoietic stem cells and divergent patterns of PU.1 expression within flk2(-) and flk2(+) common myeloid progenitors. The survey also revealed significant differences in the level of the housekeeping transcript GAPDH across the surveyed populations, which demonstrates caveats of normalizing expression data to endogenous controls and underscores the need to put gene measurement on an absolute, copy-per-cell basis.

Publishing year

2006

Language

English

Pages

17807-17812

Publication/Series

Proceedings of the National Academy of Sciences

Volume

103

Issue

47

Document type

Journal article

Publisher

National Academy of Sciences

Topic

  • Cell and Molecular Biology

Keywords

  • PU.1
  • microfluidics
  • gene profiling
  • hematopoiesis
  • stem cells

Status

Published

ISBN/ISSN/Other

  • ISSN: 1091-6490