1. Engineering dynamical control of cell fate switching using synthetic phospho-regulons
- Author
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Gordley, Russell M, Williams, Reid E, Bashor, Caleb J, Toettcher, Jared E, Yan, Shude, and Lim, Wendell A
- Subjects
Bioengineering ,Biotechnology ,Underpinning research ,1.1 Normal biological development and functioning ,Generic health relevance ,Cell Engineering ,Cell Lineage ,Gene Regulatory Networks ,Mitogen-Activated Protein Kinases ,Phosphorylation ,Regulon ,Saccharomyces cerevisiae ,Saccharomyces cerevisiae Proteins ,Synthetic Biology ,Time Factors ,Transcription ,Genetic ,dynamical control ,synthetic biology ,phosphorylation - Abstract
Many cells can sense and respond to time-varying stimuli, selectively triggering changes in cell fate only in response to inputs of a particular duration or frequency. A common motif in dynamically controlled cells is a dual-timescale regulatory network: although long-term fate decisions are ultimately controlled by a slow-timescale switch (e.g., gene expression), input signals are first processed by a fast-timescale signaling layer, which is hypothesized to filter what dynamic information is efficiently relayed downstream. Directly testing the design principles of how dual-timescale circuits control dynamic sensing, however, has been challenging, because most synthetic biology methods have focused solely on rewiring transcriptional circuits, which operate at a single slow timescale. Here, we report the development of a modular approach for flexibly engineering phosphorylation circuits using designed phospho-regulon motifs. By then linking rapid phospho-feedback with slower downstream transcription-based bistable switches, we can construct synthetic dual-timescale circuits in yeast in which the triggering dynamics and the end-state properties of the ON state can be selectively tuned. These phospho-regulon tools thus open up the possibility to engineer cells with customized dynamical control.
- Published
- 2016