EXPERIMENT_TITLE: GSM7990267: HiC_3BKO-IAA10d; Homo sapiens; Hi-C
EXPERIMENT_DESIGN_LIBRARY_DESCRIPTOR_LIBRARY_CONSTRUCTION_PROTOC: Hi-C assays were performed with an Arima Hi-C plus kit (Arima Genomics #A510008) according to manufacturer's Instructions starting from 2*106 cells/reaction. ibraries preparation were performed with a Kapa Hyper Prep Kits with KAPA Library Amplification Primer Mix (Roche, 07962347001), quantified by Qubit (Invitrogen), checked on a Tapestation device (Agilent) and Pair-End sequenced on a NovaSeq platform (Illumina).
STUDY_DESCRIPTOR_STUDY_ABSTRACT: DNA methylation (DNAme) is a key epigenetic mark that regulates critical biological processes maintaining overall genome stability. Given its pleiotropic function, studies of DNAme dynamics are crucial, but currently available tools to interfere with DNAme have limitations and major cytotoxic side effects. Here, we present cell models that allow inducible and reversible DNAme modulation through DNMT1 depletion. By dynamically assessing whole genome and locus-specific effects of induced passive demethylation through cell divisions, we reveal a cooperative activity between DNMT1 and DNMT3B, but not of DNMT3A, to maintain and control DNAme. We show that gradual loss of DNAme is accompanied by progressive and reversible changes in heterochromatin, compartmentalization, and peripheral localization. DNA methylation loss coincided with a gradual reduction of cell fitness due to G1 arrest, but with minor level of mitotic failure. Altogether, this system allows DNMTs and DNA methylation studies with fine temporal resolution, which may help to reveal the etiologic link between DNAme dysfunction and human disease. Overall design: Chromosome conformation capture (Hi-C) in DLD1 cells derived cell lines with or without DNMTs
SAMPLE_TITLE: HiC_3BKO-IAA10d
SAMPLE_SAMPLE_ATTRIBUTES_SAMPLE_ATTRIBUTE: [{"TAG": "source_name", "VALUE": "DLD1"}, {"TAG": "cell line", "VALUE": "DLD1"}, {"TAG": "cell type", "VALUE": "colorectal adenocarcinoma cell line"}, {"TAG": "genotype", "VALUE": "DNMT1-AID;DNMT3b -/-"}, {"TAG": "treatment", "VALUE": "Auxin 10 days"}, {"TAG": "geo_loc_name", "VALUE": "missing"}, {"TAG": "collection_date", "VALUE": "missing"}]
Pool_Member_@attributes_sample_title: HiC_3BKO-IAA10d
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