Genome-Wide Profiling of DNA Methyltransferases in Mammalian Cells
| Autor: | Tuncay Baubec, Massimiliano Manzo, Christina Ambrosi |
|---|---|
| Přispěvatelé: | University of Zurich, Vavouri, Tanya, Peinado, Miguel A, Baubec, Tuncay |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Methyltransferase Immunoprecipitation Protein Array Analysis ChIP Biotin Computational biology Article DNA sequencing CpG islands 03 medical and health sciences chemistry.chemical_compound 1311 Genetics seq generation sequencing 1312 Molecular Biology Animals Humans Binding site Promoter Regions Genetic DNA Modification Methylases Binding Sites biology Chemistry DNA methyltransferases DNA DNA Methylation Next Avidin 10226 Department of Molecular Mechanisms of Disease Chromatin 030104 developmental biology CpG site biology.protein 570 Life sciences Chromatin immunoprecipitation Software Genome-Wide Association Study In vivo biotinylation |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493977673 Methods Mol Biol |
| Popis: | Chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) is currently the method of choice to determine binding sites of chromatin-associated factors in a genome-wide manner. Here, we describe a method to investigate the binding preferences of mammalian DNA methyltransferases (DNMT) based on ChIP-seq using biotin-tagging. Stringent ChIP of DNMT proteins based on the strong interaction between biotin and avidin circumvents limitations arising from low antibody specificity and ensures reproducible enrichment. DNMT-bound DNA fragments are ligated to sequencing adaptors, amplified and sequenced on a high-throughput sequencing instrument. Bioinformatic analysis gives valuable information about the binding preferences of DNMTs genome-wide and around promoter regions. This method is unconventional due to the use of genetically engineered cells; however, it allows specific and reliable determination of DNMT binding. |
| Databáze: | OpenAIRE |
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