PT  - JOURNAL ARTICLE
AU  - Lauren Gambill
AU  - August Staubus
AU  - Andrea Ameruoso
AU  - James Chappell
TI  - A split ribozyme that links detection of a native RNA to orthogonal protein outputs
AID  - 10.1101/2022.01.12.476080
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.01.12.476080
4099  - http://biorxiv.org/content/early/2022/01/12/2022.01.12.476080.short
4100  - http://biorxiv.org/content/early/2022/01/12/2022.01.12.476080.full
AB  - 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 StatementThe authors have declared no competing interest.