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A dual-purpose polymerase engineered for direct sequencing of pseudouridine and queuosine

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2023

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Kaur, Navpreet
Marchand, Virginie
Motorin, Yuri
Ehrenhofer-Murray, Ann E

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Nucleic Acids Research. Oxford University Press. 2023, 51(8), pp. 3971-3987. ISSN 0305-1048. eISSN 1362-4962. Available under: doi: 10.1093/nar/gkad177

Zusammenfassung

More than 170 posttranscriptional RNA modifications are so far known on both coding and noncoding RNA species. Within this group, pseudouridine (Ψ) and queuosine (Q) represent conserved RNA modifications with fundamental functional roles in regulating translation. Current detection methods of these modifications, which both are reverse transcription (RT)-silent, are mostly based on the chemical treatment of RNA prior to analysis. To overcome the drawbacks associated with indirect detection strategies, we have engineered an RT-active DNA polymerase variant called RT-KTq I614Y that produces error RT signatures specific for Ψ or Q without prior chemical treatment of the RNA samples. Combining this polymerase with next-generation sequencing techniques allows the direct identification of Ψ and Q sites of untreated RNA samples using a single enzymatic tool.

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ISO 690HUBER, Luisa B., Navpreet KAUR, Melanie HENKEL, Virginie MARCHAND, Yuri MOTORIN, Ann E EHRENHOFER-MURRAY, Andreas MARX, 2023. A dual-purpose polymerase engineered for direct sequencing of pseudouridine and queuosine. In: Nucleic Acids Research. Oxford University Press. 2023, 51(8), pp. 3971-3987. ISSN 0305-1048. eISSN 1362-4962. Available under: doi: 10.1093/nar/gkad177
BibTex
@article{Huber2023-03-27dualp-66545,
  year={2023},
  doi={10.1093/nar/gkad177},
  title={A dual-purpose polymerase engineered for direct sequencing of pseudouridine and queuosine},
  number={8},
  volume={51},
  issn={0305-1048},
  journal={Nucleic Acids Research},
  pages={3971--3987},
  author={Huber, Luisa B. and Kaur, Navpreet and Henkel, Melanie and Marchand, Virginie and Motorin, Yuri and Ehrenhofer-Murray, Ann E and Marx, Andreas}
}
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