Abstract
Riboswitches have a number of characteristics that make them ideal regulatory elements for a wide range of synthetic biology applications. To maximize their utility, methods are required to create custom riboswitches de novo or to modify existing riboswitches to suit specific experimental needs. This chapter describes such a method, which exploits fluorescence-activated cell sorting (FACS) to quickly and efficiently sort through large libraries of riboswitch-like sequences to identify those with the desired activity. Suggestions for the experimental setup are provided, along with detailed protocols for testing and optimizing FACS conditions FACS selection steps, and follow-up assays to identify and characterize individual riboswitches.
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Acknowledgments
The riboswitch work in the Li Lab has been supported by the Natural Science and Engineering Research Council of Canada (NSERC).
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Ghazi, Z., Fowler, C.C., Li, Y. (2014). Artificial Riboswitch Selection: A FACS-Based Approach. In: Ogawa, A. (eds) Artificial Riboswitches. Methods in Molecular Biology, vol 1111. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-755-6_5
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DOI: https://doi.org/10.1007/978-1-62703-755-6_5
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