@article{Aschenbrenner2018-01-08Engin-41793,
  year={2018},
  doi={10.1002/anie.201710209},
  title={Engineering of a DNA Polymerase for Direct m<sup>6</sup>A Sequencing},
  number={2},
  volume={57},
  issn={0570-0833},
  journal={Angewandte Chemie International Edition},
  pages={417--421},
  author={Aschenbrenner, Joos and Werner, Stephan and Marchand, Virginie and Adam, Martina and Motorin, Yuri and Helm, Mark and Marx, Andreas}
}

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    <dc:rights>Attribution-NonCommercial 4.0 International</dc:rights>
    <dc:contributor>Motorin, Yuri</dc:contributor>
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    <dc:contributor>Aschenbrenner, Joos</dc:contributor>
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    <dcterms:title>Engineering of a DNA Polymerase for Direct m&lt;sup&gt;6&lt;/sup&gt;A Sequencing</dcterms:title>
    <dc:contributor>Adam, Martina</dc:contributor>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2018-03-15T08:34:42Z</dc:date>
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    <dc:contributor>Helm, Mark</dc:contributor>
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    <dc:contributor>Marx, Andreas</dc:contributor>
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    <dc:contributor>Werner, Stephan</dc:contributor>
    <dcterms:abstract xml:lang="eng">Methods for the detection of RNA modifications are of fundamental importance for advancing epitranscriptomics. N&lt;sup&gt;6&lt;/sup&gt;-methyladenosine (m&lt;sup&gt;6&lt;/sup&gt;A) is the most abundant RNA modification in mammalian mRNA and is involved in the regulation of gene expression. Current detection techniques are laborious and rely on antibody-based enrichment of m&lt;sup&gt;6&lt;/sup&gt;A-containing RNA prior to sequencing, since m&lt;sup&gt;6&lt;/sup&gt;A modifications are generally "erased" during reverse transcription (RT). To overcome the drawbacks associated with indirect detection, we aimed to generate novel DNA polymerase variants for direct m&lt;sup&gt;6&lt;/sup&gt;A sequencing. Therefore, we developed a screen to evolve an RT-active KlenTaq DNA polymerase variant that sets a mark for N&lt;sup&gt;6&lt;/sup&gt;-methylation. We identified a mutant that exhibits increased misincorporation opposite m&lt;sup&gt;6&lt;/sup&gt;A compared to unmodified A. Application of the generated DNA polymerase in next-generation sequencing allowed the identification of m&lt;sup&gt;6&lt;/sup&gt;A sites directly from the sequencing data of untreated RNA samples.</dcterms:abstract>
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