Publikation: A DNA Polymerase Variant Senses the Epigenetic Marker 5-Methylcytosine by Increased Misincorporation
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Dysregulation of DNA methylation is associated with human disease, particularly cancer, and the assessment of aberrant methylation patterns holds great promise for clinical diagnostics. However, DNA polymerases do not effectively discriminate between processing 5-methylcytosine (5 mC) and unmethylated cytosine, resulting in the silencing of methylation information during amplification or sequencing. As a result, current detection methods require multi-step DNA conversion treatments or careful analysis of sequencing data to decipher individual 5 mC bases. To overcome these challenges, we propose a novel DNA polymerase-mediated 5 mC detection approach. Here, we describe the engineering of a thermostable DNA polymerase variant derived from Thermus aquaticus with altered fidelity towards 5 mC. Using a screening-based evolutionary approach, we have identified a DNA polymerase that exhibits increased misincorporation towards 5 mC during DNA synthesis. This DNA polymerase generates mutation signatures at methylated CpG sites, allowing direct detection of 5 mC by reading an increased error rate after sequencing without prior treatment of the sample DNA.
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HENKEL, Melanie, Alexander FILLBRUNN, Virginie MARCHAND, Govindan RAGHUNATHAN, Michael R. BERTHOLD, Yuri MOTORIN, Andreas MARX, 2024. A DNA Polymerase Variant Senses the Epigenetic Marker 5-Methylcytosine by Increased Misincorporation. In: Angewandte Chemie International Edition. Wiley. 2024, 63(48), e202413304. ISSN 1433-7851. eISSN 1521-3773. Verfügbar unter: doi: 10.1002/anie.202413304BibTex
@article{Henkel2024-11-25Polym-71225, title={A DNA Polymerase Variant Senses the Epigenetic Marker 5-Methylcytosine by Increased Misincorporation}, year={2024}, doi={10.1002/anie.202413304}, number={48}, volume={63}, issn={1433-7851}, journal={Angewandte Chemie International Edition}, author={Henkel, Melanie and Fillbrunn, Alexander and Marchand, Virginie and Raghunathan, Govindan and Berthold, Michael R. and Motorin, Yuri and Marx, Andreas}, note={Article Number: e202413304} }
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