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Dissection of uniparental reproduction barriers in mice using haploid embryonic stem cells [RNA-Seq]

ABSTRACT: Dissection of uniparental reproduction barriers in mice using haploid embryonic stem cells [RNA-Seq]

PROVIDER: PRJNA491432 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR7852030_1.fastq.gz	Fastqsanger.gz
	SRR7852030_2.fastq.gz	Fastqsanger.gz
	SRR7852021_1.fastq.gz	Fastqsanger.gz
	SRR7852021_2.fastq.gz	Fastqsanger.gz
	SRR7852031_1.fastq.gz	Fastqsanger.gz

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Dissection of uniparental reproduction barriers in mice using haploid embryonic stem cells [RRBS]

Project description:Uniparental reproduction is widespread among lower vertebrates, but not in mammals. Researchers have produced bimaternal mice with a deletion of the H19 imprinted region. However, the mechanism of a single deletion in an immature oocyte sufficient to cross uniparental barriers is unknown. We found bimaternal-derived mice to be defective in multiple aspects. More importantly, bipaternal reproduction has not been achieved in mammals. Thus, the barrier of uniparental reproduction in mammals has not been elucidated. We identified a DNA hypomethylation status in haploid embryonic stem cells similar to that in primordial germ cells, with which bimaternal and bipaternal mice can be obtained by injection of haploid ES cells with genetic modifications. The phenotypic analyses of derived mice support the genetic conflict theory of genomic imprinting. Taken together, our results highlight the factors necessary for crossing uniparental reproduction barriers in mammals.

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Dissection of uniparental reproduction barriers in mice using haploid embryonic stem cells [RNA-Seq]

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Dissection of uniparental reproduction barriers in mice using haploid embryonic stem cells

Project description:Dissection of uniparental reproduction barriers in mice using haploid embryonic stem cells

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Dissection of uniparental reproduction barriers in mice using haploid embryonic stem cells [RRBS]

Project description:Dissection of uniparental reproduction barriers in mice using haploid embryonic stem cells [RRBS]

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Dissection of uniparental reproduction barriers in mice using haploid embryonic stem cells

Project description:This SuperSeries is composed of the SubSeries listed below.

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Nucleosomes are context-specific, H2A.Z modulated barriers to RNA polymerase

Project description:Nucleosomes are barriers to transcription in vitro, however, their effects on RNA polymerase in vivo are unknown. Here we describe a simple and general strategy to comprehensively map the positions of elongating and arrested RNA Polymerase II (RNAPII) at nucleotide resolution. Our results suggest that nucleosomes present significant, context-specific barriers to RNAPII in vivo that can be tuned by the incorporation of H2A.Z. 8 sets of two replicates each of paired-end and 3 sets of two replicates each of single-end samples were sequenced and analyzed.

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Mouse preimplantation imprinting [WGBS]

Project description:In mammals, chromatin marks at imprinted genes are asymmetrically inherited from each parent to control gene expression. Many genomic imprints are determined by differentially methylated regions (DMRs), but these have not been comprehensively mapped physically or functionally in mouse preimplantation embryos. To address this, we measured genome-wide DNA methylation in individual haploid uniparental parthenogenetic haploid (ph) and androgenetic haploid (ah) E3.5 blastocysts by micro-whole-genome bisulfite sequencing (µWGBS). For comparison, we also included ahESC and phESC lines in the analysis.Comparison of DNA methylomes from uniparental blastocysts with those of control blastocysts (produced by intracytoplasmic sperm injection) identified 859 DMRs. Haploid ES cells showed overall erosion of those DMRs, in contrast to diploid ES cells that relatively maintained differential methylation.

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Project description:Methylation profiles of chr12-16 were generated by meDIP and array hybridisation in 3 cases with maternal uniparental disomy of chromosome 15, and three cases of paternal uniparental disomy of chromosome 15. Comparison of these profiles reveals differentially methylated (imprinted) regions on chromosome 15.

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Epigenetic integrity of paternal imprints enhances the developmental potential of androgenetic haploid embryonic stem cells

Project description:The use of two inhibitors of Mek1/2 and Gsk3β (2i) promotes the generation of mouse diploid and haploid embryonic stem cells (ESCs) from the inner cell mass of biparental and uniparental blastocysts, respectively. However, a system enabling long-term maintenance of imprints in ESCs has proven challenging. Here, we report that usage of a two-step a2i (alternative two inhibitors of Src and Gsk3β, TSa2i) derivation/culture protocol results in the establishment of androgenetic haploid ESCs (AG-haESCs) with stable DNA methylation at paternal DMRs (differentially DNA methylated regions) up to passage 60 that can efficiently support generating mice upon oocyte injection. We also show coexistence of H3K9me3 marks and ZFP57 bindings with intact DMR methylations. Furthermore, we demonstrate that TSa2i-treated AG-haESCs are a heterogeneous cell population regarding paternal DMR methylation. Strikingly, AG-haESCs with late passages display increased paternal-DMR methylations and improved developmental potential compared to early-passage cells, in part through the enhanced proliferation of H19-DMR hypermethylated cells. Together, we establish AG-haESCs that can long-term maintain paternal imprints.

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