A suppressor tRNA-mediated feedforward loop eliminates leaky gene expression in bacteria
| dc.citation.issueNumber | 5 | |
| dc.citation.journalTitle | Nucleic Acids Research | |
| dc.citation.volumeNumber | 49 | |
| dc.contributor.author | Ho, Joanne M.L. | |
| dc.contributor.author | Miller, Corwin A. | |
| dc.contributor.author | Parks, Sydney E. | |
| dc.contributor.author | Mattia, Jacob R. | |
| dc.contributor.author | Bennett, Matthew R. | |
| dc.date.accessioned | 2021-04-21T15:46:21Z | |
| dc.date.available | 2021-04-21T15:46:21Z | |
| dc.date.issued | 2021 | |
| dc.description.abstract | Ligand-inducible genetic systems are the mainstay of synthetic biology, allowing gene expression to be controlled by the presence of a small molecule. However, ‘leaky’ gene expression in the absence of inducer remains a persistent problem. We developed a leak dampener tool that drastically reduces the leak of inducible genetic systems while retaining signal in Escherichia coli. Our system relies on a coherent feedforward loop featuring a suppressor tRNA that enables conditional readthrough of silent non-sense mutations in a regulated gene, and this approach can be applied to any ligand-inducible transcription factor. We demonstrate proof-of-principle of our system with the lactate biosensor LldR and the arabinose biosensor AraC, which displayed a 70-fold and 630-fold change in output after induction of a fluorescence reporter, respectively, without any background subtraction. Application of the tool to an arabinose-inducible mutagenesis plasmid led to a 540-fold change in its output after induction, with leak decreasing to the level of background mutagenesis. This study provides a modular tool for reducing leak and improving the fold-induction within genetic circuits, demonstrated here using two types of biosensors relevant to cancer detection and genetic engineering. | |
| dc.identifier.citation | Ho, Joanne M.L., Miller, Corwin A., Parks, Sydney E., et al.. "A suppressor tRNA-mediated feedforward loop eliminates leaky gene expression in bacteria." <i>Nucleic Acids Research,</i> 49, no. 5 (2021) Oxford University Press: https://doi.org/10.1093/nar/gkaa1179. | |
| dc.identifier.digital | gkaa1179 | |
| dc.identifier.doi | https://doi.org/10.1093/nar/gkaa1179 | |
| dc.identifier.uri | https://hdl.handle.net/1911/110302 | |
| dc.language.iso | eng | |
| dc.publisher | Oxford University Press | |
| dc.rights | This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. | |
| dc.rights.uri | http://creativecommons.org/licenses/by/4.0/), | |
| dc.title | A suppressor tRNA-mediated feedforward loop eliminates leaky gene expression in bacteria | |
| dc.type | Journal article | |
| dc.type.dcmi | Text | |
| dc.type.publication | publisher version |
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