Abstract
Abstract The FOXP3 gene is an X chromosome encoded transcription factor that is critical for the function of regulatory T (Treg) cells that maintain immunological self-tolerance and homeostasis. FOXP3 expression is primarily controlled by DNA methylation and the necessary development of Treg cells requires the establishment of Treg-specific DNA methylation patterns. FOXP3 and its epigenetic control genes have been associated with several immune-related conditions such as auto-immunity, asthma, food allergies and cancer. Here, we have conducted a study comparing the sensitivity to methylation status within our assays that cover FOXP3 using pyrosequencing, targeted bisulfite NGS (tbNGS), and methylation-sensitive high-resolution melting (MS-HRM) using a variety of different starting materials in both human and mice. As we transition away from pyrosequencing, we ensure that both tbNGS and MS-HRM result in methylation profiles which do not exhibit significant differences in sensitivity and accuracy to pyrosequencing, while also reducing the cost to the end user.
Published Version
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