Recent advances have demonstrated the use of RNA-based control devices to program sophisticated cellular functions. The ability to quantitatively tailor regulatory activities of these devices would support their broader implementation in cellular networks.We developed a high-throughput, quantitative, cell-based screening strategy to support the rapid generation of ribozyme-based RNA switches with user-specified regulatory activities. A dual reporter system was implemented to resolve device regulatory activity from cellular noise. By randomizing individual components within the ribozyme device platform, we isolated new device sequences with enhanced regulatory activities, including identifying sequences that increased catalytic activity by more than tenfold.For broader integration within biological networks, genes controlled by synthetic switches must cross the threshold activity associated with phenotypic change in the absence and presence of input signal. Our screening strategy provides a generalizable approach for quantitatively tailoring regulatory activities of genetic control elements, enabling devices to be tailored for specific systems.

High-throughput, quantitative cell-based screen for efficient tailoring of RNA device activity

244th National Meeting (2012)

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