Synthetic Cassettes for pH-Mediated Sensing, Counting, and Containment.

Cell Reports
Finn StirlingPamela A. Silver

Abstract

As pH is fundamental to all biological processes, pH-responsive bacterial genetic circuits enable precise sensing in any environment. Where the unintentional release of engineered bacteria poses a concern, coupling pH sensing to the expression of a toxin creates an effective bacterial containment system. Here, we present a pH-sensitive kill switch (acidic termination of replicating population [acidTRP]), based on the Escherichia coli asr promoter, with a survival ratio of <1 in 106. We integrate acidTRP with cryodeath to produce a 2-factor containment system with a combined survival ratio of <1 in 1011 while maintaining evolutionary stability. We further develop a pulse-counting circuit with single-cell readout for each administered stimulus pulse. We use this pulse counter to record multiple pH changes and combine it with acidTRP to make a 2-count acid-sensitive kill switch. These results demonstrate the ability to build complex genetic systems for biological containment.

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Citations

Jul 27, 2021·Current Opinion in Biomedical Engineering·Hye-In SonLingchong You
Jul 31, 2021·Nature Biomedical Engineering·Tiffany ChienTal Danino

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