ChIP-chip by array of human HeLa cells to characterize Methyl-CpG binding sites, RNA Polymerase 2 and methylated DNA regions
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
SUBMITTER: Robert D
PROVIDER: E-MEXP-2634 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:Methylated DNA binding protein 2 (MBD2) has been shown to bind specific methylated promoters and suppress transcription. Here we systematically investigate MBD2 suppression by overexpressing MBD2 in MCF-10A cells and generating genome-wide promoter methylation profiles in order to identify methylation changes likely to affect gene expression levels. MCF-10A cells were infected with MBD2 lentivirus in order to increase MBD2 expression. DNA was extracted from both infected and non-infected cells and enriched for methylated DNA through the method of methylated DNA immunoprecipitation (MeDIP). Enriched and total DNA was hybridized in different channels to 244K custom microarrays (Agilent Technologies) that contained probes covering at 100bp-spacing all transcription start sites from -800bp to 200bp (human, Ensembl v44) and all microRNAs in miRBase from 250bp before to 250bp after the microRNA. Three technical replicates were hybridized for infected and non-infected cells.
2013-06-13 | E-GEOD-47857 | biostudies-arrayexpress
Project description:We used a systems approach to identify target genes and genome wide chromatin binding preferences of the transcription factor WUSCHEL (WUS) in Arabidopsis thaliana. First, we recorded changes in the transcriptome after genetically perturbing the regulatory system of the stem cell niche at various developmental stages by loss-of-function and inducible over-expression alleles of WUS and its antagonist CLV3. Then, identified direct WUS target genes by chromatin immuno-precipitation (ChIP.
2010-06-01 | E-MEXP-2499 | biostudies-arrayexpress
Project description:Effect of insulin on the cardiomyocyte global transcriptome and RNAs recruited to polysomes. <br>For comparison with endothelin-1
2010-07-04 | E-MEXP-2527 | biostudies-arrayexpress
Project description:Extensive loss of DNA methylation is a hallmark of cancer. The role of hypomethylation in altering gene expression in cancer cells has been poorly understood. Hepatic cellular carcinoma (HCC) is one of the most common human cancers. We use HCC as a model to investigate hypomethylation in cancer by a combination of methylated DNA immunoprecipitation and hybridization with comprehensive promoter arrays. We identify approximately 2,800 promoters that are hypomethylated in tumor samples. The hypomethylated promoters appear in clusters across the genome suggesting a high-level organization behind the epigenomic changes in cancer. The genes whose promoters are demethylated are mainly involved in cell growth, cell adhesion and communication, signal transduction, mobility and invasion; functions that are essential for cancer progression and metastasis. Previous studies suggested that MBD2 was involved in demethylation of uPA and MMP2 genes in human breast and prostate cancer cell lines. We extend these results here showing that whereas MBD2 depletion in normal liver cells has little or no effect, its depletion in the human hepatocellular carcinoma cell line HepG2 and the adenocarcinoma cell line SkHep1 results in suppression of cell growth, anchorage-independent growth and invasiveness, as well as an increase in promoter methylation and silencing of several of the genes that are hypomethylated in tumors. Our studies establish for the first time the rules governing hypomethylation of promoters in liver cancer and define the potential functional role of hypomethylation in cancer. Cancerous and normal adjacent tissue samples were obtained from 11 patients with HCC from the Chinese National Human Genome Center at Shanghai, China (Dr. Ze-Guang Han). For three patients, the cancer samples were dissected using laser capture microdissection technique. All patients provided written informed consent, and the Ethics Committee from the Chinese National Human Genome Center at Shanghai approved all aspects of this study. Human HCC HepG2 cells and adenocarcinoma SkHep1 cells were purchased from ATCC (HB8065 and HTB52, respectively, USA), whereas human untransformed hepatocytes (normal hepatocytes, NorHep) were obtained from Celprogen (33003-02, USA). Purified DNA from cancerous samples and normal adjacent samples, as well as HepG2 and NorHep cells, was enriched for methylated DNA using the MeDIP protocol developed by Cedar's group (PMID 16444255). The labeled input and bound DNA samples were hybridized to a custom-designed 244K promoter tiling array that contained probes covering all transcription start sites at intervals from 800 bp upstream to 200 bp downstream of all genes described in Ensembl (version 44) and within 250 bp of the ~400 microRNAs from miRBase, all at 100 bp-spacing.
2011-06-04 | E-GEOD-29720 | biostudies-arrayexpress
Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series
2013-02-04 | E-GEOD-38242 | biostudies-arrayexpress
Project description:DNA methylation profiling of normal prostates from organ donors and prostate cancer metastases from a rapid autopsy cohort of lethal metastatic prostate cancer Multiple anatomically distinct metastases from each of five patients. Normal prostate samples from organ donors
2013-02-04 | E-GEOD-38073 | biostudies-arrayexpress
Project description:Human papillomavirus infection is the cause of essentially all cases of cervical premalignant lesions and cervical cancer. Primary screening using HPV DNA testing with cytology triage has been shown to be more sensitive and, importantly, more specific than conventional testing based on cytology among women more than 35 years of age. <br><br><br><br>The aim of the experiment was to evaluate a HPV genotyping microarray method using patient sample DNA. HPV genotyping is based on multiplex PCR (PGMY-t primers) followed by ligation step where two probes are ligated together if a matching template is present in the mixture. The ligated probes are then detected on microarray.
2011-12-10 | E-MEXP-3466 | biostudies-arrayexpress
Project description:This SuperSeries is composed of the following subset Series: GSE29720: The landscape of promoter DNA hypomethylation in liver cancer (MeDIP-chip data) GSE29721: The landscape of promoter DNA hypomethylation in liver cancer (expression data) Refer to individual Series
2011-06-04 | E-GEOD-29722 | biostudies-arrayexpress
Project description:The aim of the study was to use microarray for profiling the microbiota in anaerobic digestion process. The probes are ssDNA molecules that are ligated into circular molecules if a complementary target sequence is present in the sample DNA. Ligated probes are PCR amplified with a labeled primer, and the amplicons are hybridized on DNA microarray by tag sequences.
2012-02-06 | E-MEXP-3541 | biostudies-arrayexpress
Project description:Chronic physical aggression (CPA) is characterized by frequent use of physical aggression from early childhood to adolescence. Observed in approximately 5% of males, it is associated with early childhood adverse environments. DNA methylation, a covalent modification of DNA, regulates genome function and has been associated with early childhood adverse environments. We analyzed T cell DNA genome-wide promoter methylation profiles from two groups of adult males assessed annually for frequency of physical aggression between 6 and 15 years of age: a group with CPA and a control group. Whole genome methylation analysis was performed using MeDIP followed by hybridization to microarrays. We mapped the promoter regions of 20 000 genes and 400 microRNAs. The subjects were recruited from participants in two longitudinal studies of child development9,47. We recruited two groups of Caucasian males who were born in families with a low socioeconomic status and were living at the time of the present study within 200km from our laboratory. The first group had a history of high physical aggression from age 6 to 15 years (chronic physical aggression group, CPA). The second group was recruited from the same longitudinal studies but included only those who did not have a history of high physical aggression from age 6 to 15 (Control group). A total of 65 eligible subjects agreed to participate (8 CPA and 57 controls). Using custom-designed microarrays with 244K probes tiling the promoter regions of 20000 genes and 400 microRNAs, we obtained DNA methylation profiles by meDIP-chip. Each profile was generated in triplicate.
2013-09-30 | E-GEOD-50674 | biostudies-arrayexpress