Objectif Acquired immunity to foreign pathogens depends on functional B and T cells. The objective of this proposal is to elucidate the transcriptional control of lymphocyte development at three stages by deciphering the transcriptional networks specifying pro-B and pro-T cells in early lymphopoiesis and plasma cells in terminal B cell differentiation.To this end, we generated knock-in mice carrying a biotin acceptor sequence at the C-terminus of transcription factors, which can be biotinylated by transgenic co-expression of the E. coli biotin ligase BirA. In vivo biotinylation facilitates antibody-independent precipitation of these transcription factors by streptavidin pulldown (Bio-ChIP). Preliminary Bio-ChIP sequencing experiments validated this approach for genome-wide identification of transcription factor target genes.Bio-ChIP sequencing will be used to identify the target genes of key transcription factors controlling the development of pro-B cells (Ikaros, E2A, STAT5, EBF1, Pax5, PU.1 IRF4), pro-T cells (Notch1, RBP-J, GATA3, Ikaros, E2A, STAT5) and plasma cells (Blimp1, IRF4, XBP1). RNA sequencing of wild-type and mutant lymphocytes will determine the regulated target genes of these factors, which are ultimately relevant for the elucidation of transcriptional networks. The function of selected target genes at central nodes of these networks will be analyzed by the latest miR30-shRNA knockdown technology. Finally, regulatory complexes interacting with these transcription factors will be identified by streptavidin-pulldown purification and mass spectrometry followed by their integration into the transcriptional networks by ChIP-seq mapping to the transcription factor target genes.These experiments will provide fundamentally new molecular insight into the generation of all three lymphocyte stages and will contribute to a better understanding of how deregulation of the transcriptional control promotes the development of lymphoid malignancies. Champ scientifique natural sciencesbiological sciencesgeneticsRNAnatural scienceschemical sciencesanalytical chemistrymass spectrometry Programme(s) FP7-IDEAS-ERC - Specific programme: "Ideas" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013) Thème(s) ERC-AG-LS6 - ERC Advanced Grant - Immunity and infection Appel à propositions ERC-2011-ADG_20110310 Voir d’autres projets de cet appel Régime de financement ERC-AG - ERC Advanced Grant Institution d’accueil FORSCHUNGSINSTITUT FUR MOLEKULARE PATHOLOGIE GESELLSCHAFT MBH Contribution de l’UE € 2 499 305,00 Adresse CAMPUS-VIENNA-BIOCENTER 1 1030 Wien Autriche Voir sur la carte Région Ostösterreich Wien Wien Type d’activité Private for-profit entities (excluding Higher or Secondary Education Establishments) Contact administratif Tanja Winkler (Ms.) Chercheur principal Meinrad Busslinger (Prof.) Liens Contacter l’organisation Opens in new window Site web Opens in new window Coût total Aucune donnée Bénéficiaires (1) Trier par ordre alphabétique Trier par contribution de l’UE Tout développer Tout réduire FORSCHUNGSINSTITUT FUR MOLEKULARE PATHOLOGIE GESELLSCHAFT MBH Autriche Contribution de l’UE € 2 499 305,00 Adresse CAMPUS-VIENNA-BIOCENTER 1 1030 Wien Voir sur la carte Région Ostösterreich Wien Wien Type d’activité Private for-profit entities (excluding Higher or Secondary Education Establishments) Contact administratif Tanja Winkler (Ms.) Chercheur principal Meinrad Busslinger (Prof.) Liens Contacter l’organisation Opens in new window Site web Opens in new window Coût total Aucune donnée