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The SBE7 enhancer is active in Shh expressing brain tissue at E10.5

ABSTRACT: Using ATAC-seq, we identified an accessible chromatin region within the Shh expressing cell population of eGFP/Cre-Shh mice, which did not correspond to SBE2-4. This open region displayed high levels of vertebrate conservation, and did not map to a known CTCF site. Further analysis revealed the coordinates of this presumptive active enhancer to correspond to the recently identified SBE7 enhancer.

ORGANISM(S): Mus musculus

PROVIDER: GSE158074 | GEO | 2020/09/17

REPOSITORIES: GEO

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The SBE7 enhancer is active in Shh expressing brain tissue at E10.5

Project description:The SBE7 enhancer is active in Shh expressing brain tissue at E10.5

| PRJNA663892 | ENA

H3K27ac in WT and Shh-/-, and Shh-/-;Gli3-/- E10.5 limb buds

Project description:Purpose: The response to Hedgehog signaling in the limb is driven by GLI bound enhancers. While most Hh targets in the developing limb bud appear to be regulated by the activity of GLI-repressor, the role of GLI activator in this regulation is unclear. Here we assess the roles of GLI activator and repressor in regulating H3K27ac to activate their enhancers. Methods: ChIP-seq was performed in Embryonic day 10.5 mouse limb buds from WT, Shh-/- and Shh-/-Gli3-/- embryos Results: We found that most GLI binding regions can be activated in the absence of both GLI activator and repressor, while a small subset require GLI activator for activity. These indicate that most GLI enhancers in the limb are regulated by GLI repression.

2020-01-21 | GSE133712 | GEO

Shh and ZRS enhancer co-localization is specific to the zone of polarizing activity

Project description:Limb-specific expression of Shh is regulated by the long-range (~one megabasepair distant) ZRS enhancer. In the mouse, murine limb bud restricted spatiotemporal expression of Shh occurs from ~E10 until E11.5 at the distal posterior margin is essential for the correct formation of the autopod. Here, we have analyzed the higher-order chromatin conformation of Shh in expressing and non-expressing tissues, both by fluorescence in situ hybridization (FISH) and by chromosome conformation capture (5C). Conventional and super-resolution light microscopy identified significantly elevated frequences of Shh/ZRS co-localization only in the Shh expressing regions of the limb bud consistent with the formation of an enhancer-promoter loop. However, Shh-ZRS spatial distances were consistently shorter than intervening distances to a neural enhancer in all tissues and developmental stages analyzed – regardless of Shh expression. 5C also identified a topologically associating domain (TAD) over the Shh-ZRS genomic region and enriched interactions between Shh and ZRS, but in the head, body and limb buds of E11.5 embryos, so also not linked to Shh expression. We show that gene-enhancer (Shh/ZRS) co-localization correlates with the spatiotemporal domain of limb bud-specific Shh expression, but that close Shh/ZRS proximity in the nucleus occurs regardless of whether the gene or enhancer is active. We suggest that this constrained chromatin configuration optimises the opportunity for the active enhancer to locate and instigate Shh expression.

2016-05-03 | GSE79947 | GEO

H3K27ac in WT and Shh-/-, and Shh-/-;Gli3-/- E10.5 limb buds

Project description:H3K27ac in WT and Shh-/-, and Shh-/-;Gli3-/- E10.5 limb buds

| PRJNA552304 | ENA

Chromatin conformation of the Shh locus in limbs.

Project description:Expression of Shh in the limb is dependent on a distant enhancer located 850kb away from the gene promoter. We performed chromatin conformation capture (4C-Seq) analysis of the Shh locus in E11.5 mouse limbs so as to better understand the relationship between this regulatory interaction and the structural organisation of the locus. The experiments were performed on wild-type animals as well as on animals carrying an engineered chromosomal inversion (INV(6-C2) (INV6)) . The different viewpoints used (Shh, ZRS, Nom1, Rbm33, Rnf32) enable to characterize the 3D organisation of the locus in the different contexts. The inversion INV6 disrupts the normal topological organisation of the Shh locus in a compact 3D-domain and prevents the physical and regulatory interaction between Shh and its limb enhancer, even though the genomic distance separating the two elements is reduced compared to WT.

2017-01-12 | E-MTAB-4980 | biostudies-arrayexpress

RNA sequencing of mouse littermate wild-type and Shh null E10.5 forelimbs [Illumina]

Project description:We wanted to identify differentially expressed genes in wild-type and Shh null E10.5 mouse forelimbs Used two wild-type technical replicates and two Shh null technical replicates UT-Genome and Analysis Facility

2015-07-01 | E-GEOD-58645 | biostudies-arrayexpress

Transcriptome of wild type and Shh limb buds at E10.5

Project description:The transcriptome of mouse limb buds of Shh mutant embryos was compared to the transcriptome of limb buds of wild type embryos at embryonic day E10.5

2011-05-01 | E-MEXP-3142 | biostudies-arrayexpress

RNA sequencing of mouse littermate wild-type and Shh null E10.5 forelimbs [Illumina]

Project description:We wanted to identify differentially expressed genes in wild-type and Shh null E10.5 mouse forelimbs

2015-07-01 | GSE58645 | GEO

Cis-regulatory architecture of a brain-signaling center predates the origin of chordates

Project description:Genomic approaches have predicted hundreds of thousands of tissue specific cis-regulatory sequences, but the determinants critical to their function and evolutionary history are mostly unknown1-4. Here, we systematically decode a set of brain enhancers active in the zona limitans intrathalamica (zli), a signaling center essential for vertebrate forebrain development via the secreted morphogen, Sonic hedgehog (Shh)5,6. We apply a de novo motif analysis tool to identify six position-independent sequence motifs together with their cognate transcription factors that are essential for zli enhancer activity and Shh expression in the mouse embryo. Using knowledge of this regulatory lexicon, we discover novel Shh zli enhancers in mice, and a functionally equivalent element in hemichordates, indicating an ancient origin of the Shh zli regulatory network that predates the chordate phylum. These findings establish a paradigm for delineating functionally conserved enhancers in the absence of overt sequence homologies, and over extensive evolutionary distances. Gene expression profiles from the mouse zona limitans intrathalamica (ZLI) region at E10.5

2016-03-25 | E-GEOD-78005 | biostudies-arrayexpress

Developmentally regulated Shh expression is robust to TAD perturbations

Project description:Topologically Associating Domains (TADs) have been proposed to both guide and constrain enhancer activity. Shh is located within a TAD known to contain all its enhancers. To investigate the importance of chromatin conformation and TAD integrity on developmental gene regulation, we have manipulated the Shh TAD – creating internal deletions, deleting CTCF sites, and deleting and inverting sequences at TAD boundaries. Chromosome conformation capture and fluorescence in situ hybridisation assays were used to investigate the changes in chromatin conformation that result from these manipulations. Our data suggest that these substantial alterations in TAD structure has no readily detectable effect on Shh expression patterns during development – except where enhancers are deleted - and results in no detectable phenotypes. Only in the case of a larger deletion of one TAD boundary could ectopic influence of the Shh limb enhancer be detected on a gene (Mnx1) in the neighbouring TAD. Our data suggests that, contrary to expectations, the developmental regulation of Shh expression is remarkably robust to TAD perturbations.

2019-08-15 | GSE135840 | GEO

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