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PMID: 25417166 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

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

Cell ·Vol. 159 ·No. 4 ·2014-11-06 ·Pages 925-39

Green AA, Silver PA, Collins JJ, Yin P

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.

MeSH Terms
Computer Simulation Escherichia coli/genetics,metabolism Gene Expression Regulation Gene Regulatory Networks RNA/chemistry Regulatory Sequences, Ribonucleic Acid Synthetic Biology
Chemicals
Regulatory Sequences, Ribonucleic Acid RNA
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Green Alexander A
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA.
Silver Pamela A
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA; Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.
Collins James J
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA; Howard Hughes Medical Institute, Department of Biomedical Engineering and Center of Synthetic Biology, Boston University, Boston, MA 02215, USA.
Yin Peng
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA; Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA. Electronic address: py@hms.harvard.edu.
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Article Info
Journal
Cell
Abbr.
Cell
ISSN
1097-4172
DOI
10.1016/j.cell.2014.10.002
Email
py@hms.harvard.edu
Published
2014-11-06
Epub
2014-00-23
Pages
925-39
Language
English
Region
United States
NLM ID
0413066
PMCID
PMC4265554
Subset
IM
Grants
NIH HHS · 1DP2OD007292 · United States
NIBIB NIH HHS · 1R01EB018659 · United States
Howard Hughes Medical Institute · United States
NIH HHS · DP2 OD007292 · United States
NIBIB NIH HHS · R01 EB018659 · United States
Corrections
CommentIn
PMID: 25519125
External Links
PubMed source DOI: 10.1016/j.cell.2014.10.002 PMC Full Text View journal details
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