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Modulating transcription through development of semi-synthetic yeast core promoters
Altering gene expression regulation by promoter engineering is a very effective way to fine-tune heterologous pathways in eukaryotic hosts. Typically, pathway building approaches in yeast still use a limited set of long, native promoters. With the today’s introduction of longer and more complex path...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6830820/ https://www.ncbi.nlm.nih.gov/pubmed/31689317 http://dx.doi.org/10.1371/journal.pone.0224476 |
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author | Decoene, Thomas De Maeseneire, Sofie L. De Mey, Marjan |
author_facet | Decoene, Thomas De Maeseneire, Sofie L. De Mey, Marjan |
author_sort | Decoene, Thomas |
collection | PubMed |
description | Altering gene expression regulation by promoter engineering is a very effective way to fine-tune heterologous pathways in eukaryotic hosts. Typically, pathway building approaches in yeast still use a limited set of long, native promoters. With the today’s introduction of longer and more complex pathways, an expansion of this synthetic biology toolbox is necessary. In this study we elucidated the core promoter structure of the well-characterized yeast TEF1 promoter and determined the minimal length needed for sufficient protein expression. Furthermore, this minimal core promoter sequence was used for the creation of a promoter library covering different expression strengths. This resulted in a group of short, 69 bp promoters with an 8.0-fold expression range. One exemplar had a two and four times higher expression compared to the native CYC1 and ADH1 promoter, respectively. Additionally, as it was described that the protein expression range could be broadened by upstream activating sequences (UASs), we integrated earlier described single and multiple short, synthetic UASs in front of the strongest yeast core promoter. This approach resulted to further variation in protein expression and an overall promoter library spanning a 20-fold activity range and covering a length from 69 bp to maximally 129 bp. Furthermore, the robustness of this library was assessed on three alternative carbon sources besides glucose. As such, the suitability of short yeast core promoters for metabolic engineering applications on different media, either in an individual context or combined with UAS elements, was demonstrated. |
format | Online Article Text |
id | pubmed-6830820 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-68308202019-11-14 Modulating transcription through development of semi-synthetic yeast core promoters Decoene, Thomas De Maeseneire, Sofie L. De Mey, Marjan PLoS One Research Article Altering gene expression regulation by promoter engineering is a very effective way to fine-tune heterologous pathways in eukaryotic hosts. Typically, pathway building approaches in yeast still use a limited set of long, native promoters. With the today’s introduction of longer and more complex pathways, an expansion of this synthetic biology toolbox is necessary. In this study we elucidated the core promoter structure of the well-characterized yeast TEF1 promoter and determined the minimal length needed for sufficient protein expression. Furthermore, this minimal core promoter sequence was used for the creation of a promoter library covering different expression strengths. This resulted in a group of short, 69 bp promoters with an 8.0-fold expression range. One exemplar had a two and four times higher expression compared to the native CYC1 and ADH1 promoter, respectively. Additionally, as it was described that the protein expression range could be broadened by upstream activating sequences (UASs), we integrated earlier described single and multiple short, synthetic UASs in front of the strongest yeast core promoter. This approach resulted to further variation in protein expression and an overall promoter library spanning a 20-fold activity range and covering a length from 69 bp to maximally 129 bp. Furthermore, the robustness of this library was assessed on three alternative carbon sources besides glucose. As such, the suitability of short yeast core promoters for metabolic engineering applications on different media, either in an individual context or combined with UAS elements, was demonstrated. Public Library of Science 2019-11-05 /pmc/articles/PMC6830820/ /pubmed/31689317 http://dx.doi.org/10.1371/journal.pone.0224476 Text en © 2019 Decoene et al http://creativecommons.org/licenses/by/4.0/ 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 use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Decoene, Thomas De Maeseneire, Sofie L. De Mey, Marjan Modulating transcription through development of semi-synthetic yeast core promoters |
title | Modulating transcription through development of semi-synthetic yeast core promoters |
title_full | Modulating transcription through development of semi-synthetic yeast core promoters |
title_fullStr | Modulating transcription through development of semi-synthetic yeast core promoters |
title_full_unstemmed | Modulating transcription through development of semi-synthetic yeast core promoters |
title_short | Modulating transcription through development of semi-synthetic yeast core promoters |
title_sort | modulating transcription through development of semi-synthetic yeast core promoters |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6830820/ https://www.ncbi.nlm.nih.gov/pubmed/31689317 http://dx.doi.org/10.1371/journal.pone.0224476 |
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