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A split ribozyme that links detection of a native RNA to orthogonal protein outputs

View ORCID ProfileLauren Gambill, View ORCID ProfileAugust Staubus, View ORCID ProfileAndrea Ameruoso, View ORCID ProfileJames Chappell
doi: https://doi.org/10.1101/2022.01.12.476080
Lauren Gambill
1PhD Program in Systems, Synthetic, and Physical Biology, Rice University, Houston, TX, USA 77005
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August Staubus
2Department of Biosciences, Rice University, Houston, TX, USA 77005
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Andrea Ameruoso
2Department of Biosciences, Rice University, Houston, TX, USA 77005
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James Chappell
1PhD Program in Systems, Synthetic, and Physical Biology, Rice University, Houston, TX, USA 77005
2Department of Biosciences, Rice University, Houston, TX, USA 77005
3Department of Bioengineering, Rice University, Houston, TX, USA 77005
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  • For correspondence: jc125@rice.edu
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ABSTRACT

Individual RNA remains a challenging signal to synthetically transduce into different types of cellular information. Here, we describe Ribozyme-ENabled Detection of RNA (RENDR), a plug-and-play strategy that uses cellular transcripts to template the assembly of split ribozymes, triggering splicing reactions that generate orthogonal protein outputs. To identify split ribozymes that require templating for splicing, we used laboratory evolution to evaluate the activities of different split variants of the Tetrahymena thermophila ribozyme. The best design delivered a 93-fold dynamic range of splicing with RENDR controlling fluorescent protein production in response to an RNA input. We resolved a thermodynamic model to guide RENDR design, showed how input signals can be transduced into diverse visual, chemical, and regulatory outputs, and used RENDR to detect an antibiotic resistance phenotype in bacteria. This work shows how transcriptional signals can be monitored in situ using RNA synthetic biology and converted into different types of biochemical information.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted January 12, 2022.
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A split ribozyme that links detection of a native RNA to orthogonal protein outputs
Lauren Gambill, August Staubus, Andrea Ameruoso, James Chappell
bioRxiv 2022.01.12.476080; doi: https://doi.org/10.1101/2022.01.12.476080
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A split ribozyme that links detection of a native RNA to orthogonal protein outputs
Lauren Gambill, August Staubus, Andrea Ameruoso, James Chappell
bioRxiv 2022.01.12.476080; doi: https://doi.org/10.1101/2022.01.12.476080

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