Abstract
RNAs that are transcribed and self-assemble within living cells are valuable tools for regulating and organizing cellular activities. Riboregulators, in particular, have been widely used for modulating translation and transcription in response to cognate transactivating or trigger RNAs, enabling cells to evaluate logic operations and to respond to environmental cues. Herein we detail a set of methods for the rapid construction and testing of prokaryotic riboregulators in Escherichia coli. These methods enable construction of dozens of riboregulator plasmids at the same time without the use of restriction enzymes. Furthermore, they facilitate rapid screening of devices and can be applied to a variety of other self-assembling in vivo RNA systems.
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Acknowledgment
This work was supported by start-up funds from Arizona State University.
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Green, A.A. (2017). Construction and In Vivo Testing of Prokaryotic Riboregulators. In: Bindewald, E., Shapiro, B. (eds) RNA Nanostructures . Methods in Molecular Biology, vol 1632. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7138-1_19
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DOI: https://doi.org/10.1007/978-1-4939-7138-1_19
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