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

Warren, Luigi and Bryder, David and Weissman, Irving L. and Quake, Stephen R. (2006) Transcription factor profiling in individual hematopoietic progenitors by digital RT-PCR. Proceedings of the National Academy of Sciences of the United States of America, 103 (47). pp. 17807-17812. ISSN 0027-8424. https://resolver.caltech.edu/CaltechAUTHORS:20090917-153509721

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Abstract

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.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1073/pnas.0608512103DOIUNSPECIFIED
http://www.pnas.org/content/103/47/17807.abstractPublisherUNSPECIFIED
Additional Information:Copyright ©2006 by the National Academy of Sciences. Contributed by Irving L. Weissman, September 26, 2006 (received for review July 2, 2006). Published online before print November 10, 2006, doi: 10.1073/pnas.0608512103 This work was supported by U.S. Public Health Service Grant 5 P01 DK 053074 (from the National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health) (I.L.W. and D.B.), National Science Foundation Grant 0535870 (S.R.Q. and L.W.), and the National Institutes of Health Director's Pioneer Award (S.R.Q. and L.W.). Author contributions: L.W., D.B., and S.R.Q. designed research; L.W. and D.B. performed research; L.W., D.B., S.R.Q., and I.L.W. analyzed data; and L.W., D.B., S.R.Q., and I.L.W. wrote the paper. Conflict of interest statement: S.Q. is a founder, shareholder, and consultant for Fluidigm Corporation.
Funders:
Funding AgencyGrant Number
National Institute of Diabetes and Digestive and Kidney Diseases5 P01 DK 053074
NSF0535870
NIHUNSPECIFIED
Subject Keywords:gene profiling; hematopoiesis; microfluidics; PU.1; stem cells
Issue or Number:47
Record Number:CaltechAUTHORS:20090917-153509721
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20090917-153509721
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:15933
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:02 Oct 2009 16:48
Last Modified:03 Oct 2019 01:05

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