Journal
METHODS
Volume 187, Issue -, Pages 13-27Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2020.07.006
Keywords
Next-generation sequencing; Target bisulfite-seq; DNA methylation; Biomarker discovery; Epigenetic dock
Funding
- National Institutes of Health [T32CA201160]
- UCLA QCBio Collaboratory Fellowship
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Cytosine methylation, a well-studied epigenetic modification, plays crucial roles in development and disease, with patterns varying among cells and changing with age. DNA methylation arrays have been developed to detect methylation status in a cost-effective way, while a Targeted Bisulfite Sequencing method offers a promising approach for biomarker discovery across different phenotypes.
Cytosine methylation is one of the best studied epigenetic modifications. In mammals, DNA methylation patterns vary among cells and is mainly found in the CpG context. DNA methylation is involved in important processes during development and differentiation and its dysregulation can lead to or is associated with diseases, such as cancer, loss-of-imprinting syndromes and neurological disorders. It has been also shown that DNA methylation at the cellular, tissue and organism level varies with age. To overcome the costs of Whole-Genome Bisulfite Sequencing, the gold standard method to detect 5-methylcytosines at a single base resolution, DNA methylation arrays have been developed and extensively used. This method allows one to assess the status of a fraction of the CpG sites present in the genome of an organism. In order to combine the relatively low cost of Methylation Arrays and digital signals of bisulfite sequencing, we developed a Targeted Bisulfite Sequencing method that can be applied to biomarker discovery for virtually any phenotype. Here we describe a comprehensive step-by-step protocol to build a DNA methylation-based epigenetic clock.
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