Genomics

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Transcription factor ChIP-seq in expanded human hematopoietic stem and progenitor cells


ABSTRACT: While many individual transcription factors are known to regulate hematopoietic differentiation, major aspects of the global architecture of hematopoiesis remain unknown. Here, we profiled gene expression in 38 distinct purified populations of human hematopoietic cells and used probabilistic models of gene expression and analysis of cis-elements in gene promoters to decipher the general organization of their regulatory circuitry. We identified modules of highly co-expressed genes, some of which are restricted to a single lineage, but most are expressed at variable levels across multiple lineages. We found densely interconnected cis-regulatory circuits and a large number of transcription factors that are differentially expressed across hematopoietic states, suggesting a more complex regulatory system than previously assumed. We functionally validated a subset of candidate factors using RNA interference and ChIP-Seq. Our dataset, analytic tools, and findings, available on a web-based portal, provide a unique resource to study the regulatory architecture of hematopoiesis. Human CD133-positive Umbilical Cord Blood (UCB) cells were cross-linked with formaldehyde for 20 min. DNA was enriched by chromatin immunoprecipitation (ChIP) and analyzed by Solexa sequencing. A sample of whole cell extract (WCE) was sequenced and used as the background to determine enrichment. ChIP was performed using an antibody against total TAL1 (Santa Cruz SC-12984), PU.1/SPI1 (Santa Cruz SC-352X), IKAROS (Santa Cruz SC-9859X) and MEIS1 (Abcam ab19867). This submission represents the ChIP-seq component of the study.

ORGANISM(S): Homo sapiens  

SUBMITTER: Lee N Lawton   Richard A Young 

PROVIDER: E-GEOD-26014 | ArrayExpress | 2011-01-26

SECONDARY ACCESSION(S): GSE26014SRP005406PRJNA135475

REPOSITORIES: GEO, ArrayExpress, ENA

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Though many individual transcription factors are known to regulate hematopoietic differentiation, major aspects of the global architecture of hematopoiesis remain unknown. Here, we profiled gene expression in 38 distinct purified populations of human hematopoietic cells and used probabilistic models of gene expression and analysis of cis-elements in gene promoters to decipher the general organization of their regulatory circuitry. We identified modules of highly coexpressed genes, some of which  ...[more]

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