Published August 10, 2021 | Version v1

Riboswitch-inspired toehold riboregulators for gene regulation in Escherichia coli

Description

This dataset comprises flow cytometry data accompanying a publication on the development of synthetic riboregulators.

These riboregulators were inspired by the architecture of naturally occurring riboswitches and toehold-mediated strand displacement. Specifically, we adopt the toehold switch hairpin and inserted regulatory sequences within the loop region of which accessibility can be controlled by toehold-mediated strand displacement. We utilized this design principle to develop toehold translation repressor and toehold transcriptional repressor, which regulate mCherry expression in E. coli in translational and transcriptional levels with certain ON/OFF ratios. Furthermore, we combined these two riboregulators and developed them into a NOR gate switch that can regulate downstream GFP expression in E. coli with different input conditions of trigger RNA. We used flow cytometry to quantify the expression level of the NOR gate switch under different inputs.

Notes

Flow cytometry of riboregulator logic gates was performed using a BD FACSMelodyTM cell sorter. Flow cytometry data were analyzed using FlowJo 10.1r1 software (FLOWJO).

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