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Engineering ligand-responsive RNA controllers in yeast through the assembly of RNase III tuning modules
The programming of cellular networks to achieve new biological functions depends on the development of genetic tools that link the presence of a molecular signal to gene-regulatory activity. Recently, a set of engineered RNA controllers was described that enabled predictable tuning of gene expressio...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3130268/ https://www.ncbi.nlm.nih.gov/pubmed/21355039 http://dx.doi.org/10.1093/nar/gkr090 |
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author | Babiskin, Andrew H. Smolke, Christina D. |
author_facet | Babiskin, Andrew H. Smolke, Christina D. |
author_sort | Babiskin, Andrew H. |
collection | PubMed |
description | The programming of cellular networks to achieve new biological functions depends on the development of genetic tools that link the presence of a molecular signal to gene-regulatory activity. Recently, a set of engineered RNA controllers was described that enabled predictable tuning of gene expression in the yeast Saccharomyces cerevisiae through directed cleavage of transcripts by an RNase III enzyme, Rnt1p. Here, we describe a strategy for building a new class of RNA sensing-actuation devices based on direct integration of RNA aptamers into a region of the Rnt1p hairpin that modulates Rnt1p cleavage rates. We demonstrate that ligand binding to the integrated aptamer domain is associated with a structural change sufficient to inhibit Rnt1p processing. Three tuning strategies based on the incorporation of different functional modules into the Rnt1p switch platform were demonstrated to optimize switch dynamics and ligand responsiveness. We further demonstrated that these tuning modules can be implemented combinatorially in a predictable manner to further improve the regulatory response properties of the switch. The modularity and tunability of the Rnt1p switch platform will allow for rapid optimization and tailoring of this gene control device, thus providing a useful tool for the design of complex genetic networks in yeast. |
format | Online Article Text |
id | pubmed-3130268 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-31302682011-07-06 Engineering ligand-responsive RNA controllers in yeast through the assembly of RNase III tuning modules Babiskin, Andrew H. Smolke, Christina D. Nucleic Acids Res Synthetic Biology and Chemistry The programming of cellular networks to achieve new biological functions depends on the development of genetic tools that link the presence of a molecular signal to gene-regulatory activity. Recently, a set of engineered RNA controllers was described that enabled predictable tuning of gene expression in the yeast Saccharomyces cerevisiae through directed cleavage of transcripts by an RNase III enzyme, Rnt1p. Here, we describe a strategy for building a new class of RNA sensing-actuation devices based on direct integration of RNA aptamers into a region of the Rnt1p hairpin that modulates Rnt1p cleavage rates. We demonstrate that ligand binding to the integrated aptamer domain is associated with a structural change sufficient to inhibit Rnt1p processing. Three tuning strategies based on the incorporation of different functional modules into the Rnt1p switch platform were demonstrated to optimize switch dynamics and ligand responsiveness. We further demonstrated that these tuning modules can be implemented combinatorially in a predictable manner to further improve the regulatory response properties of the switch. The modularity and tunability of the Rnt1p switch platform will allow for rapid optimization and tailoring of this gene control device, thus providing a useful tool for the design of complex genetic networks in yeast. Oxford University Press 2011-07 2011-02-25 /pmc/articles/PMC3130268/ /pubmed/21355039 http://dx.doi.org/10.1093/nar/gkr090 Text en © The Author(s) 2011. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Synthetic Biology and Chemistry Babiskin, Andrew H. Smolke, Christina D. Engineering ligand-responsive RNA controllers in yeast through the assembly of RNase III tuning modules |
title | Engineering ligand-responsive RNA controllers in yeast through the assembly of RNase III tuning modules |
title_full | Engineering ligand-responsive RNA controllers in yeast through the assembly of RNase III tuning modules |
title_fullStr | Engineering ligand-responsive RNA controllers in yeast through the assembly of RNase III tuning modules |
title_full_unstemmed | Engineering ligand-responsive RNA controllers in yeast through the assembly of RNase III tuning modules |
title_short | Engineering ligand-responsive RNA controllers in yeast through the assembly of RNase III tuning modules |
title_sort | engineering ligand-responsive rna controllers in yeast through the assembly of rnase iii tuning modules |
topic | Synthetic Biology and Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3130268/ https://www.ncbi.nlm.nih.gov/pubmed/21355039 http://dx.doi.org/10.1093/nar/gkr090 |
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