Toehold switches: de-novo-designed regulators of gene expression

Efforts to construct synthetic networks in living cells have been hindered by the limited number of regulatory components that provide wide dynamic range and low crosstalk. Here, we report a class of de-novo-designed prokaryotic riboregulators called toehold switches that activate gene expression in...

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Bibliographic Details
Published in:Cell 2014-11, Vol.159 (4), p.925-939
Main Authors: Green, Alexander A, Silver, Pamela A, Collins, James J, Yin, Peng
Format: Article
Language:English
Subjects:
Computer Simulation
Escherichia coli - genetics
Escherichia coli - metabolism
Gene Expression Regulation
Gene Regulatory Networks
Regulatory Sequences, Ribonucleic Acid
RNA - chemistry
Synthetic Biology
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Summary:Efforts to construct synthetic networks in living cells have been hindered by the limited number of regulatory components that provide wide dynamic range and low crosstalk. Here, we report a class of de-novo-designed prokaryotic riboregulators called toehold switches that activate gene expression in response to cognate RNAs with arbitrary sequences. Toehold switches provide a high level of orthogonality and can be forward engineered to provide average dynamic range above 400. We show that switches can be integrated into the genome to regulate endogenous genes and use them as sensors that respond to endogenous RNAs. We exploit the orthogonality of toehold switches to regulate 12 genes independently and to construct a genetic circuit that evaluates 4-input AND logic. Toehold switches, with their wide dynamic range, orthogonality, and programmability, represent a versatile and powerful platform for regulation of translation, offering diverse applications in molecular biology, synthetic biology, and biotechnology.
ISSN:0092-8674
1097-4172
DOI:10.1016/j.cell.2014.10.002