Simultaneous quantification of rDNA methylation and copy number: Constraints to natural variation in humans and cell lines.

ABSTRACT: Ribosomal DNA (rDNA) copy number (CN) varies widely both across and within species. In humans, it has been hypothesized that rDNA transcriptional activity is partially regulated by rDNA methylation status. Here we describe a rapid and scalable approach for the simultaneous detection of methylated and unmethylated rDNA copy number using a well characterized digital PCR technology. The linear range for detecting rDNA CN and methylation is from 0.3 ng to 20 ng DNA with a practical range from 1.25 to 15 ng DNA and can be normalized with a single copy gene, ACTB, to reflect copies per genome. Our approach detects 28S rDNA methylation using a modified methyl-specific/bisulfite PCR targeting a region with at least four CpGs. In a population of healthy donors, we observed wide variation in both me

SUBMITTER: Smith SF 

PROVIDER: S-EPMC12614545 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature