A general design strategy for protein-responsive riboswitches in mammalian cells

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2014
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Ausländer, Simon
Stücheli, Pascal
Ausländer, David
Fussenegger, Martin
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Nature Methods. 2014, 11, pp. 1154-1160. ISSN 1548-7091. eISSN 1548-7105. Available under: doi: 10.1038/nmeth.3136
Zusammenfassung

RNAs are ideal for the design of gene switches that can monitor and program cellular behavior because of their high modularity and predictable structure-function relationship. We have assembled an expression platform with an embedded modular ribozyme scaffold that correlates self-cleavage activity of designer ribozymes with transgene translation in bacteria and mammalian cells. A design approach devised to screen ribozyme libraries in bacteria and validate variants with functional tertiary stem-loop structures in mammalian cells resulted in a designer ribozyme with a protein-binding nutR-boxB stem II and a selected matching stem I. In a mammalian expression context, this designer ribozyme exhibited dose-dependent translation control by the N-peptide, had rapid induction kinetics and could be combined with classic small molecule–responsive transcription control modalities to construct complex, programmable genetic circuits.

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540 Chemie
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Riboswitches, Synthetic biology, Gene regulation
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ISO 690AUSLÄNDER, Simon, Pascal STÜCHELI, Charlotte REHM, David AUSLÄNDER, Jörg S. HARTIG, Martin FUSSENEGGER, 2014. A general design strategy for protein-responsive riboswitches in mammalian cells. In: Nature Methods. 2014, 11, pp. 1154-1160. ISSN 1548-7091. eISSN 1548-7105. Available under: doi: 10.1038/nmeth.3136
BibTex
@article{Auslander2014gener-30093,
  year={2014},
  doi={10.1038/nmeth.3136},
  title={A general design strategy for protein-responsive riboswitches in mammalian cells},
  volume={11},
  issn={1548-7091},
  journal={Nature Methods},
  pages={1154--1160},
  author={Ausländer, Simon and Stücheli, Pascal and Rehm, Charlotte and Ausländer, David and Hartig, Jörg S. and Fussenegger, Martin}
}
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