Transcription factor profiling in individual hematopoietic progenitors by digital RT-PCR.
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.
Warren
Luigi
L
Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA.
Bryder
David
D
Weissman
Irving L
IL
Quake
Stephen R
SR
D000818:Animals; D020869:Gene Expression Profiling / Q000295:instrumentation / Q000379:methods; D018507:Gene Expression Regulation, Developmental; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D014157:Transcription Factors