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Differentiation of human cardiomyocyte and thymic epithelial cells from H1 control hESCs and H1 hESCs uniquely modified to be DNA methylated at the promoter CpG island of MLH1.

ABSTRACT: Differentiation of human cardiomyocyte and thymic epithelial cells from H1 control hESCs and H1 hESCs uniquely modified to be DNA methylated at the promoter CpG island of MLH1.

PROVIDER: PRJNA863661 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR20715944.fastq.gz	Fastqsanger.gz
	SRR20715945.fastq.gz	Fastqsanger.gz
	SRR20715946.fastq.gz	Fastqsanger.gz
	SRR20715947.fastq.gz	Fastqsanger.gz
	SRR20715948.fastq.gz	Fastqsanger.gz

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Differentiation of human cardiomyocyte and thymic epithelial cells from H1 control hESCs and H1 hESCs uniquely modified to be DNA methylated at the promoter CpG island of MLH1.

Project description:Engineering Pluripotent DNA Methylation by CpG Island Methylation Response (CIMR) to Synthetic CpG-free ssDNA Insertion Cellular differentiation requires global changes to DNA methylation (DNAme), where it functions to regulate transcription factor and chromatin remodeling activity, and genome interpretation. Here, we describe a simple DNAme engineering approach in pluripotent stem cells (PSCs), extending across large stretches of CpG dense “islands (CGIs).” Integration of synthetic CpG free single-stranded DNA (ssDNA) induces a target CpG Island Methylation Response (CIMR) in multiple PSC lines, Nt2d1 embryonal carcinoma cells, and mouse PSCs, but not in highly methylated CpG Island Methylator Phenotype (CIMP) positive cancer lines. CIMR DNAme at MLH1 spans the CGI, is robustly maintained throughout cellular differentiation, suppresses target gene activity, and sensitizes derived cardiomyocytes and thymic epithelial cells to the chemotherapy cisplatin. Additional CIMR DNAme is reported on at TP53 and ONECUT1 CGIs. Collectively, this new resource enables total CpG Island DNAme engineering in pluripotency and the genesis of novel epigenetic models of development and disease

2023-06-21 | GSE210107 | GEO

RNA Sequencing of H1 WT hESCs, H1 QSER1 KO hESCs, H1 TET1 KO hESCs, H1 QSER1/TET1 DKO hESCs, WT Day10 EBs, QSER1 KO Day10 EBs, TET1 KO Day10 EBs, QSER1/TET1 DKO Day10 EBs, WT PP1, QSER1 KO PP1, TET1 KO PP1, and QSER1/TET1 DKO PP1.

Project description:RNA Sequencing of H1 WT hESCs, H1 QSER1 KO hESCs, H1 TET1 KO hESCs, H1 QSER1/TET1 DKO hESCs, WT Day10 embryoid bodies (EBs), QSER1 KO Day10 EBs, TET1 KO Day10 EBs, QSER1/TET1 DKO Day10 EBs, WT pancreatic progenitors (PP1), QSER1 KO PP1, TET1 KO PP1, and QSER1/TET1 DKO PP1. DNA methylation is essential to mammalian development, and dysregulation can cause serious pathological conditions. Key enzymes responsible for deposition and removal of DNA methylation are known, but how they cooperate to tightly regulate the methylation landscape remains a central question. Utilizing a knockin DNA methylation reporter, we performed a genome-wide CRISPR/Cas screen in human embryonic stem cells to discover DNA methylation regulators. The top screen hit was an uncharacterized gene QSER1, which proved to be a key guardian of bivalent promoters and poised enhancers of developmental genes, especially those residing in DNA methylation valleys (or canyons). We further demonstrate cooperation of QSER1 and TET1 through genetic and biochemical interactions to inhibit DNMT3-mediated de novo methylation and safeguard developmental programs.

2021-03-19 | GSE169209 | GEO

5-hydroxymethylcytosine (5hmC) profiling of H1 WT hESCs

Project description:5-hydroxymethylcytosine (5hmC) profiling of H1 WT hESCs

| PRJNA630896 | ENA

Project description:RNA Sequencing of H1 WT hESCs, H1 QSER1 KO hESCs, H1 TET1 KO hESCs, H1 QSER1/TET1 DKO hESCs, WT Day10 EBs, QSER1 KO Day10 EBs, TET1 KO Day10 EBs, QSER1/TET1 DKO Day10 EBs, WT PP1, QSER1 KO PP1, TET1 KO PP1, and QSER1/TET1 DKO PP1.

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RNA-seq of H1 (WA-01) hESCs cultured in 3 conditions (mTeSR1, 4iLA + feeders and FINE)

Project description:RNA-seq was performed to look at global differences in gene expression between human embryonic stem cells (hESCs) particularly H1 cell line (WA-01, passage 23-40) cultured in distinct conditions. In details we compared cells adapted to primed (mTeSR1) and two naïve states: feeder dependent (4iLA + feeder) and feeder free (FINE) states.

2019-11-01 | E-MTAB-8216 | biostudies-arrayexpress

Methyl capture sequencing (Truseq Epic from Illumina) of H1 WT, H1 QSER1 KO, H1 TET1 KO, H1 DNMT3B KO, H1 QSER1 KOpm (passaged matched with DKOs), H1 QSER1/TET1 DKO, and H1 QSER1/DNMT3B DKO hESCs

Project description:Methyl capture sequencing (Truseq Epic from Illumina) of H1 WT, H1 QSER1 KO, H1 TET1 KO, H1 DNMT3B KO, H1 QSER1 KOpm (passaged matched with DKOs), H1 QSER1/TET1 DKO, and H1 QSER1/DNMT3B DKO hESCs

| PRJNA715660 | ENA

Primary constitutional MLH1 epimutations: a focal epigenetic event [FFPE data sets]

Project description:Background: Constitutional MLH1 epimutations are characterized by monoallelic methylation of the MLH1 promoter throughout normal tissues, accompanied by allele-specific silencing. The mechanism underlying primary MLH1 epimutations is currently unknown. The aim of this study was to perform an in-depth characterization of constitutional MLH1 epimutations targeting the aberrantly methylated region around MLH1 and other genomic loci. Methods: Twelve MLH1 epimutation carriers, 61 Lynch syndrome patients and 41 healthy controls, were analyzed by Infinium Human Methylation 450K beadchip, and targeted molecular techniques were used to characterize the MLH1 epimutation in carriers and their inheritance pattern. Results: No nucleotide or structural variants were identified in-cis on the epimutated allele in ten carriers, in which intergenerational methylation erasure was demonstrated in two, suggesting primary type of epimutation. CNVs outside the MLH1 locus were found in two cases. EPM2AIP1-MLH1 CpG island was identified as the sole differentially methylated region in MLH1 epimutation carriers compared to controls. Conclusion: Primary constitutional MLH1 epimutations arise as a focal epigenetic event at the EPM2AIP1-MLH1 CpG island in the absence of cis-acting genetic variants. Further molecular characterization is needed to elucidate the mechanistic basis of MLH1 epimutations and their heritability/reversibility.

2017-11-28 | GSE107352 | GEO

Primary constitutional MLH1 epimutations: a focal epigenetic event [blood data sets]

2017-11-28 | GSE107351 | GEO

RNA-seq and ChIP-seq studies of cardiomyocyte differentiation from H1 hESCs

Project description:RNA-seq and ChIP-seq studies of cardiomyocyte differentiation from H1 hESCs

| PRJNA777795 | ENA

Ssu72 and Ser5P-RNAPII ChIPseq in hESCs. GROseq analysis in siCtrl and siSSu72 transfected hESCs.

Project description:Ssu72 is a Ser5P-RNAPII phosphatase, nevertheless few targets have been characterized and its role in mammal cells remains elusive. Using ChIPseq and GROseq technologies we aimed to identify Ssu72 targets in human embryonic stem cells. For the ChIPseq experiments, H1 hESCs growing in mTeSR media were doubled fixed with DSG 2mM (45min) and FA 1% (20min) and processed to chromatin immunoprecipitation. For GROseq experiments, H1 hESCs were transfected with siCtrl or siSsu72 siRNAs using RNAimax (Invitrogene) for 48h. Then, run-on experiment was done with isolated nuclei. Nascent RNA purification and subsequent steps needed before sequencing were carried out essentially as described in Core et al, 2008, Science 319(5871): 1791-1792.

2014-11-01 | E-GEOD-61393 | biostudies-arrayexpress

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