Extended-representation bisulfite sequencing of gene regulatory elements in multiplexed samples and single cells
العنوان: | Extended-representation bisulfite sequencing of gene regulatory elements in multiplexed samples and single cells |
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المؤلفون: | Peter van Galen, Ayush T. Raman, Martin J. Aryee, Volker Hovestadt, Sarah J. Shareef, Samantha M. Bevill, Bradley E. Bernstein |
المصدر: | Nat Biotechnol |
بيانات النشر: | Springer Science and Business Media LLC, 2021. |
سنة النشر: | 2021 |
مصطلحات موضوعية: | Bisulfite sequencing, Biomedical Engineering, Bioengineering, Computational biology, Biology, Applied Microbiology and Biotechnology, Genome, Article, Histones, 03 medical and health sciences, 0302 clinical medicine, Humans, Sulfites, Gene Regulatory Networks, Epigenetics, Copy-number variation, Gene, 030304 developmental biology, 0303 health sciences, High-Throughput Nucleotide Sequencing, Methylation, DNA Methylation, CpG site, DNA methylation, Molecular Medicine, CpG Islands, Single-Cell Analysis, 030217 neurology & neurosurgery, Biotechnology |
الوصف: | The biological roles of DNA methylation have been elucidated by profiling methods based on whole-genome or reduced-representation bisulfite sequencing, but these approaches do not efficiently survey the vast numbers of noncoding regulatory elements in mammalian genomes. Here we present a extended representation bisulfite sequencing (XRBS) method for targeted profiling of DNA methylation. Our design strikes a balance between expanding coverage of regulatory elements and reproducibly enriching informative CpG dinucleotides in promoters, enhancers, and CTCF binding sites. Barcoded DNA fragments are pooled prior to bisulfite conversion, allowing multiplex processing and technical consistency in low input samples. Application of XRBS to single leukemia cells enabled us to evaluate genetic copy-number variations and methylation variability across individual cells. Our analysis highlights heterochromatic H3K9me3 regions as having the highest cell-to-cell variability in their methylation, likely reflecting inherent epigenetic instability of these late replicating regions, compounded by differences in cell cycle stages among sampled cells. |
تدمد: | 1546-1696 1087-0156 |
URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d724943e6ee822e3fcd6581583db03a7 https://doi.org/10.1038/s41587-021-00910-x |
حقوق: | OPEN |
رقم الأكسشن: | edsair.doi.dedup.....d724943e6ee822e3fcd6581583db03a7 |
قاعدة البيانات: | OpenAIRE |
تدمد: | 15461696 10870156 |
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