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Homo sapiens

ABSTRACT: Mapping of 5-hydroxymethylcytosine by selective chemical labeling (SCL-seq) in human naive B cells and in vitro differentiated plasmablasts (P1 cells) [HTS]

PROVIDER: PRJNA293878 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR2183000.fastq.gz	Fastqsanger.gz
	SRR2183001.fastq.gz	Fastqsanger.gz
	SRR2182992.fastq.gz	Fastqsanger.gz
	SRR2182993.fastq.gz	Fastqsanger.gz
	SRR2182994.fastq.gz	Fastqsanger.gz

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Mapping of 5-hydroxymethylcytosine by selective chemical labeling (SCL-seq) in human naive B cells and in vitro differentiated plasmablasts (P1 cells) [HTS]

Project description:We report the genome-wide mapping of 5hmC in naive B cells and in vitro differentiated plasmablasts (P1 cells). In addition, H3K4me1 and H3K27ac histne marks were mapped in in vitro differentiated plasmablasts from two healthy donors (BN358 and BN369) by ChIP-seq. Genomic DNA from human primary cells was extracted and submitted to the SCL protocol using the Hydroxymethyl Collector kit from Active Motif. After sequencing on an Illumina HiSeq 2000, reads were mapped to the human hg19 genome. H3K4me1 and H3K27ac were mapped in in vitro differentiated plasmablasts (P1 cells) by ChIP-seq.

2015-10-30 | E-GEOD-72360 | biostudies-arrayexpress

Mapping of 5-hydroxymethylcytosine by selective chemical labeling (SCL-seq) in human naive B cells and in vitro differentiated plasmablasts (P1 cells) [HTS]

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Genome-wide identification of 5hmCpGs in multiple myeloma

Project description:We report the genomic localization of 5-hydroxymethylcytosines in multiple myeloma patients through the use of selective chemical labeling (SCL) and exonuclease digestion. Data highlighted super-enhancers and enhancers specifically undergoing active DNA demethylation in multiple myeloma cells.

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Mus musculus

Project description:Selective chemical labeling reveals intragenic enrichment of 5-hydroxymethylcytosine in mouse cerebellum

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Single-CpG resolution mapping of 5-hydroxymethylcytosine by chemical labeling and exonuclease digestion (SCL-exo)

Project description:We report the setup of a new method to map 5-hydroxymethylcytosine (5hmC) genome-wide at CpG resolution. The method combines selective chemical labeling by 5hmC b-glucosyltransferase and exonuclease digestion of the DNA molecules bound to streptavidin beads after biotinylation of the 5-glucosylmethylcytosines. Associated with a straightforward bioinformatic analysis, this new procedure provides a cost-effective and fast method for mapping 5hmC at high resolution.

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A highly sensitive and robust method for genome-wide 5hmC profiling of rare cell populations

Project description:Investigations of 5-hydroxymethylcytosine (5hmC) in biologically and clinically samples and models with low cell numbers have been hampered by the low sensitivity and reproducibility using current 5hmC mapping approaches. Here, we develop a selective 5hmC chemical labeling approach using tagmentation-based library preparation in order to profile nanogram levels of 5hmC isolated from ~1,000 cells (nano-hmC-Seal). Using this technology, we profiled the dynamics of 5hmC across different stages of mouse hematopoietic differentiation. Additionally, applying nano-hmC-Seal to the hematopoietic multipotent progenitor cells in an acute myeloid leukemia (AML) mouse model, we identified leukemia-specific, differentially hydroxymethylated regions that harbor previously reported and as-yet-unidentified functionally relevant factors. The change of 5hmC patterns in AML strongly correlates with the altered gene expression on a global scale. Together, our new approach offers a highly sensitive and robust method to study and detect DNA methylation dynamics from in vivo model and clinical samples. Selective 5hmC chemical labeling approach using tagmentation-based library preparation in order to profile nanogram levels of 5hmC isolated from ~1,000 cells

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Project description:Active DNA demethylation in mammals involves TET-mediated oxidation of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxycytosine (5caC). However, genome-wide detection of 5fC at single-base resolution remains challenging. Here we present a bisulfite-free method for the whole-genome analysis of 5fC, based on a selective chemical labeling of 5fC and subsequent C-to-T transition during PCR. Base-resolution 5fC maps reveal limited overlap with 5hmC, with 5fC-marked regions more active than 5hmC-marked ones.

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