title: A Brn2-Zic1 axis specifies the neuronal fate of retinoic-acid-treated embryonic stem cells.
abstract: Mouse embryonic stem cells (ESCs) treated with all-trans retinoic acid differentiate into a homogenous population of glutamatergic neurons. Although differentiation is initiated through activation of target genes by the retinoic acid receptors, the downstream transcription factors specifying neuronal fate are less well characterised. Here, we show that the transcription factor Brn2 (also known as Pou3f2) is essential for the neuronal differentiation programme. By integrating results from RNA-seq following Brn2 silencing with results from Brn2 ChIP-seq, we identify a set of Brn2 target genes required for the neurogenic programme. Further integration of Brn2 ChIP-seq data from retinoic-acid-treated ESCs and P19 cells with data from ESCs differentiated into neuronal precursors by Fgf2 treatment and that from fibroblasts trans-differentiated into neurons by ectopic Brn2 expression showed that Brn2 occupied a distinct but overlapping set of genomic loci in these differing conditions. However, a set of common binding sites and target genes defined the core of the Brn2-regulated neuronal programme, among which was that encoding the transcription factor Zic1. Small hairpin RNA (shRNA)-mediated silencing of Zic1 prevented ESCs from differentiating into neuronal precursors, thus defining a hierarchical Brn2-Zic1 axis that is essential to specify neural fate in retinoic-acid-treated ESCs.
authors.affiliation: Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDS, 1 Rue Laurent Fries, Illkirch, Cédex 67404, France.
authors.forename: Sylvia
authors.initials: S
authors.lastname: Urban
authors.affiliation: Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDS, 1 Rue Laurent Fries, Illkirch, Cédex 67404, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Kobi
authors.affiliation: Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDS, 1 Rue Laurent Fries, Illkirch, Cédex 67404, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Ennen
authors.affiliation: Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDS, 1 Rue Laurent Fries, Illkirch, Cédex 67404, France.
authors.forename: Diana
authors.initials: D
authors.lastname: Langer
authors.affiliation: Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDS, 1 Rue Laurent Fries, Illkirch, Cédex 67404, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Le Gras
authors.affiliation: Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDS, 1 Rue Laurent Fries, Illkirch, Cédex 67404, France.
authors.forename: Tao
authors.initials: T
authors.lastname: Ye
authors.affiliation: Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDS, 1 Rue Laurent Fries, Illkirch, Cédex 67404, France Equipe Labellisée of the Ligue Nationale Contre le Cancer, CNRS/INSERM/UDS, 1 Rue Laurent Fries, Illkirch, Cédex 67404, France irwin@igbmc.fr.
authors.forename: Irwin
authors.initials: I
authors.lastname: Davidson
mesh_terms: D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects*; D058732:Embryoid Bodies / Q000166:cytology / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D016678:Genome; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D050809:POU Domain Factors / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology*
chemical_list: D009419:Nerve Tissue Proteins; D050809:POU Domain Factors; D014157:Transcription Factors; C492122:Zic1 protein, mouse; C074066:Pou3f2 protein, mouse; D014212:Tretinoin
keywords: Ascl1; ChIP-seq; Fgf2; Reprogramming; Sox2