@article{Strobel2020-02-05Hight-49314,
  year={2020},
  doi={10.1038/s41467-020-14491-x},
  title={High-throughput identification of synthetic riboswitches by barcode-free amplicon-sequencing in human cells},
  number={1},
  volume={11},
  journal={Nature Communications},
  author={Strobel, Benjamin and Spöring, Maike and Klein, Holger and Blazevic, Dragica and Rust, Werner and Sayols, Sergi and Hartig, Jörg S. and Kreuz, Sebastian},
  note={Article Number: 714}
}

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    <dc:creator>Spöring, Maike</dc:creator>
    <dc:contributor>Klein, Holger</dc:contributor>
    <dc:creator>Sayols, Sergi</dc:creator>
    <dc:creator>Blazevic, Dragica</dc:creator>
    <dc:contributor>Rust, Werner</dc:contributor>
    <dc:contributor>Hartig, Jörg S.</dc:contributor>
    <dc:contributor>Kreuz, Sebastian</dc:contributor>
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    <dcterms:title>High-throughput identification of synthetic riboswitches by barcode-free amplicon-sequencing in human cells</dcterms:title>
    <dc:contributor>Blazevic, Dragica</dc:contributor>
    <dc:creator>Strobel, Benjamin</dc:creator>
    <dc:creator>Hartig, Jörg S.</dc:creator>
    <dc:language>eng</dc:language>
    <dcterms:issued>2020-02-05</dcterms:issued>
    <dcterms:abstract xml:lang="eng">Synthetic riboswitches mediating ligand-dependent RNA cleavage or splicing-modulation represent elegant tools to control gene expression in various applications, including next-generation gene therapy. However, due to the limited understanding of context-dependent structure–function relationships, the identification of functional riboswitches requires large-scale-screening of aptamer-effector-domain designs, which is hampered by the lack of suitable cellular high-throughput methods. Here we describe a fast and broadly applicable method to functionally screen complex riboswitch libraries (~1.8 × 10&lt;sup&gt;4&lt;/sup&gt; constructs) by cDNA-amplicon-sequencing in transiently transfected and stimulated human cells. The self-barcoding nature of each construct enables quantification of differential mRNA levels without additional pre-selection or cDNA-manipulation steps. We apply this method to engineer tetracycline- and guanine-responsive ON- and OFF-switches based on hammerhead, hepatitis-delta-virus and Twister ribozymes as well as U1-snRNP polyadenylation-dependent RNA devices. In summary, our method enables fast and efficient high-throughput riboswitch identification, thereby overcoming a major hurdle in the development cascade for therapeutically applicable gene switches.</dcterms:abstract>
    <dc:contributor>Spöring, Maike</dc:contributor>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2020-04-28T08:25:28Z</dc:date>
    <dc:contributor>Strobel, Benjamin</dc:contributor>
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