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Methylation is one of the essential epigenetic modifications to the DNA, which is responsible for the precise regulation of genes required for stable development and differentiation of different tissue types. Dysregulation of this process is often the hallmark of various diseases like cancer. Here, we outline one of the recent sequencing techniques, Methyl-Binding DNA Capture sequencing (MBDCap-seq), used to quantify methylation in various normal and disease tissues for large patient cohorts. We describe a detailed protocol of this affinity enrichment approach along with a bioinformatics pipeline to achieve optimal quantification. This technique has been used to sequence hundreds of patients across various cancer types as a part of the 1,000 methylome project (Cancer Methylome System).
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http://dx.doi.org/10.3791/54131 | DOI Listing |
Brain Commun
August 2025
Baker Heart and Diabetes Institute, Epigenetics in Human Health and Disease Program, Melbourne, Victoria 3004, Australia.
Focal Cortical Dysplasia (FCD) is a common cause of drug-resistant epilepsy. These abnormalities arise during embryonic development and are challenging to classify due to their complex nature. The most recent classification update of FCD incorporates genetic and epigenetic results with other clinical data for the management of epilepsy associated with these lesions.
View Article and Find Full Text PDFJ Invest Dermatol
July 2025
Department of Pathology, GROW - Research Institute for Oncology and Reproduction, Maastricht University Medical Center, Maastricht, The Netherlands. Electronic address:
Accurate risk assessment of local recurrences or metastases is essential for melanoma treatment, but current clinical parameters are suboptimal. Therefore, we conducted a comprehensive DNA methylation analysis to identify prognostic markers that could improve risk prediction in patients with melanoma. We integrated methyl-binding domain sequencing, RNA sequencing, Infinium HumanMethylation450 analyses, and The Cancer Genome Atlas data to identify potential prognostic DNA methylation markers.
View Article and Find Full Text PDFCancers (Basel)
July 2025
Department of Pathology and Laboratory Medicine, University of California, Irvine, CA 92697, USA.
: Prostate cancer (PCa) is diagnosed at an earlier median age, more advanced stage, and has worse clinical outcomes in African American (AA) men compared to European Americans (EA). : To investigate the role of aberrant DNA methylation in tumor-adjacent stroma (TAS), methyl binding domain sequencing (MBD-seq) was performed on AA ( = 17) and EA ( = 15) PCa patients. This was independently confirmed using the long interspersed nuclear element-1 (LINE-1) assay.
View Article and Find Full Text PDFAm J Physiol Cell Physiol
September 2025
Department of Pediatrics, College of Human Medicine, Michigan State University, Grand Rapids, Michigan, United States.
There is increasing evidence that the methyl-binding domain (MBD) is a protein-protein interaction motif that can function independently of methylated DNA binding. The MBD proteins found throughout plants and invertebrates duplicated into multiple vertebrate DNA and non-DNA-binding members (MBD1, MBD2, MBD3, MBD4, MBD5, MBD6, MECP2, BAZ2A, BAZ2B, SETDB1, and SETDB2). Although many invertebrate species possess MBD proteins that can bind and recognize DNA methylation, the DNA-binding function has been independently lost multiple times, with only minor alterations to the protein interaction residues.
View Article and Find Full Text PDFBiochem Biophys Res Commun
September 2025
Amity Institute of Biotechnology, Amity University, Kolkata, W.B., India. Electronic address:
Methyl-CpG-binding proteins are crucial epigenetic regulators associated with inborn genetic and neurodevelopmental disorders such as Rett Syndrome, Autism Spectrum Disorder, and Angelman Syndrome, as well as various malignancies, including colorectal, prostate, brain, breast, and endometrial cancers. The proteins that belong to Methyl Binding Domain (MBD) superfamily, have eleven members: SETDB1, SETDB2, MECP2, MBD1-6, BAZ2A, and BAZ2B. This research utilized both sequence-based and structure-based methodologies to ascertain the most detrimental mutation in MBD proteins.
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