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CHARACTERIZATION OF THE B-CELL-SPECIFIC GENE REGULATORY NETWORK BY GENOME-WIDE IDENTIFICATION OF DIRECT PAX5 TARGET GENES

Objetivo

A complex network of transcription factor interactions controls the process by which B lymphocytes develop from hematopoietic stem cells (HSCs). Pax5 has been identified as the critical transcription factor controlling B cell commitment throughout B lymphopoiesis. These analyses revealed that Pax5 commits early hematopoietic progenitors to the B cell pathway by repressing B-lineage-inappropriate genes and by simultaneously activating B-cell-specific genes. By comprehensive cDNA microarray analysis, 110 Pax5-repressed and 170 Pax5-activated genes have been identified, demonstrating that Pax5 controls a complex regulatory network involved in B cell signaling, adhesion, migration and immune function. The goal of the project proposed here will be to systematically analyze all direct Pax5 target genes in the entire genome during B cell development. Recently, a mouse strain carrying a biotin tag at the C-terminus of Pax5 has been generated. We will use this mouse strain for genome-wide identification of Pax5-binding sites by chromatin precipitation and Solexa 1G ultrahigh-throughput DNA sequencing. Islands of chromatin modifications have been shown to provide an efficient way of identifying active and repressed promoters and cis-regulatory elements. Here we propose to use chromatin immunoprecipitation (ChIP)-sequencing to map the entire mouse genome for Pax5-controlled promoters and cis-regulatory elements. Integration of this information with the identification of Pax5-binding sites will provide us with a genome-wide map of functional Pax5-binding sites, from which a list of all direct target genes of Pax5 during B cell development will be obtained. The project proposed here will constitute an essential piece of a larger group effort, which aims at elucidating the gene regulatory network interactions controlling B cell development.

Convocatoria de propuestas

FP7-PEOPLE-2007-2-1-IEF
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Coordinador

FORSCHUNGSINSTITUT FUR MOLEKULARE PATHOLOGIE GESELLSCHAFT MBH
Aportación de la UE
€ 160 749,69
Dirección
CAMPUS-VIENNA-BIOCENTER 1
1030 Wien
Austria

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Región
Ostösterreich Wien Wien
Tipo de actividad
Private for-profit entities (excluding Higher or Secondary Education Establishments)
Contacto administrativo
Harald Isemann (Mr.)
Enlaces
Coste total
Sin datos