2024 Biomarkers of Aging Conference

The predominant cytosine modification in DNA is methylcytosine (5mC) This base appears to exert a profound impact on gene expression patterns and is commonly linked to transcriptional repression. The introduction of hydroxymethylation (5hmC) adds a new layer of complexity to the conventional comprehension of DNA methylation dynamics, which is often thought to correlate with gene expression. Constrained to measuring four states of information, existing NGS- basedtechnologies sacrifice genetic information for modification calling without distinguishing between these two important modification states. Genetic and methylation data combined together presents a unique opportunity to measure these cytosine modifications against gene expression. duet multiomics solution evoC is a new sequencing technology that resolves all four genetic bases alongside the ability to distinguish modification status of cytosines, discriminating 5mC from 5hmC (6 base calling). The technology consists of pre- sequencing library prep with enzymatic conversion of DNA together with an analysis pipeline, achieving base resolution of genetics and epigenetics at high accuracy. Here we present the method, alongside a high-depth sequencing analysis of mouse embryonic stem cells. We demonstrate the potential of epigenetic modifications encoded in DNA to predict other important potential modulators of gene expression such as open chromatin and enhancer states as well as RNA expression itself.