Toehold switches

De-novo-designed regulators of gene expression

Alexander Green, Pamela A. Silver, James J. Collins, Peng Yin

Research output: Contribution to journalArticle

230 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)925-939
Number of pages15
JournalCell
Volume159
Issue number4
DOIs
StatePublished - Nov 6 2014
Externally publishedYes

Fingerprint

Regulator Genes
Gene expression
Switches
Switch Genes
RNA
Synthetic Biology
Gene Expression
Biotechnology
Genes
Molecular Biology
Genome
Molecular biology
Crosstalk
Cells
Networks (circuits)
Sensors

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Toehold switches : De-novo-designed regulators of gene expression. / Green, Alexander; Silver, Pamela A.; Collins, James J.; Yin, Peng.

In: Cell, Vol. 159, No. 4, 06.11.2014, p. 925-939.

Research output: Contribution to journalArticle

Green, Alexander ; Silver, Pamela A. ; Collins, James J. ; Yin, Peng. / Toehold switches : De-novo-designed regulators of gene expression. In: Cell. 2014 ; Vol. 159, No. 4. pp. 925-939.
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