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Hi-C of WT E14 mouse ES cells grown in serum and 2i media

ABSTRACT: Hi-C of WT E14 mouse ES cells grown in serum and 2i media

PROVIDER: PRJNA511678 | ENA |

REPOSITORIES: ENA

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Hi-C of WT E14 mouse ES cells grown in serum and 2i media

Project description:Dynamic reprogramming of global DNA methylation impacts on the genomic deposition of the Polycomb-mediated repressive histone mark H3K27me3. DNA hypomethylation in ground state embryonic stem cells (ESCs) results in a reversible redistribution of H3K27me3 from its normal target loci. Thus, a signalling induced shift of ESCs to ground state results in both DNA methylation and Polycomb patterns that are quite distinct from their primed counterparts. Here we investigated the impact of DNA methylation directed Polycomb redistribution on higher-order chromatin structure in the ground state. Using a targeted single-locus approach (FISH) we can demonstrate local decompaction at Hox loci in the ground state, which is consistent with genome-wide data (Hi-C) indicating that chromatin structure is globally altered in ground state relative to primed ESCs. Polycomb targets are similarly decompacted in hypomethylated E3.5 mouse blastocysts. ESC lines which maintain a high level of DNA methylation in ground state show no decompaction at Hox loci. Our results suggest that DNA-methylation mediated reprogramming of Polycomb binding drives higher order chromatin organisation in stem cells and early development.

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Trim24 ChIP-seq in embryonic stem (ES) cells grown by 2i+LIF (2iL) and serum media

Project description:ChIP followed by next generation sequencing against Trim24 was performed in mouse embryonic stem (ES) grown either in 2i+LIF (2iL) or serum media. Two biological replicates for each condition were sequenced using Illumina HiSeq. Two input controls (no IP) were also generated for each condition.

2016-11-08 | E-MTAB-3802 | biostudies-arrayexpress

Trim24 ChIP-seq in embryonic stem (ES) cells grown by 2i+LIF (2iL) and serum media

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Distinct DNA methylomes of 2i and serum mouse ES cells are interconvertible and reflect pre- and post-implantation embryonic states

Project description:This study describes the DNA methylation profiling using whole-genome bisulfite sequencing of mouse ES cells, either derived and maintained in 2i serum-free NDiff medium, or in the presence of serum and LIF, or maintained and derived in the presence of serum and LIF and subsequently adapted to 2i serum-free NDiff medium, or maintained and derived in the presence of 2i and LIF and subsequently adapted to 2i serum. DNA methylation profiling using whole-genome bisulfite sequencing of 14 samples, 3 different lines (E14, XT67E1, Rex/GFP-2i) of pluripotent mouse ES cells as well during conversion from 2i to serum and vice versa.

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RNA-seq analysis of ES cell populations with different functional potentials in serum/LIF and 2i/LIF, as well as analysis of the transcriptional effect of LIF in ground state (2i) culture conditions.

Project description:We report that ES cells cultured in ground state (2i and 2i/LIF) culture conditions are heterogeneous and show heterogeneus expression of extraembryonic markers. Using a highly sensitive reporter for the endoderm marker Hex we can sort Hex high and low populations from either serum/LIF or 2i/LIF and demonstrate that they have different functional properties. Here we explored the transcriptional basis of these functional differences and noted that Hex low (HV-) and Hex high (HV+) populations showed more distinct expression profiles in 2i/LIF than in serum/LIF. Additionally in 2i/LIF the HV+ population showed an upregulation of extraembryonic markers (such as trophoblast stem cell specific genes) and also imprinted genes compared to the HV- population, which is not observed when these populations are sorted from serum/LIF. We also analysed the transcriptional effect of LIF in 2i by analysing unsorted ES cells cultured in either 2i alone or 2i with LIF. We observed that the addition of LIF led to an upregulation of extraembryonic markers but did not effect the expression of pluripotency genes, other than Klf4. Additionally, the most significantly upregulated genes from 2i/LIF cultured ES cells compared to 2i cultured ES cells showed the greatest correlation to placental tissue when compared to the GNF tissue specific expression database. This analysis, alongside functional experiments, suggested that HV+ ES cells in 2i/LIF corresponded to an extraembryonically primed population of cells and that the addition of LIF supported this population.

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Quantitative multiplexed ChIP-Seq comparing mouse embryonic stem cells grown in Serum or 2i condition.

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total RNA-seq from ES-E14 (ENCSR000CWC)

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