Development of a chromosomally integrated metabolite-inducible Leu3p-alpha-IPM "off-on" gene switch.
<h4>Background</h4>Present technology uses mostly chimeric proteins as regulators and hormones or antibiotics as signals to induce spatial and temporal gene expression.<h4>Methodology/principal findings</h4>Here, we show that a chromosomally integrated yeast 'Leu3p-alpha...
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Public Library of Science (PLoS)
2010-08-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0012488&type=printable |
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| author | Maria Poulou Donald Bell Kostas Bozonelos Maria Alexiou Anthony Gavalas Robin Lovell-Badge Eumorphia Remboutsika |
| author_facet | Maria Poulou Donald Bell Kostas Bozonelos Maria Alexiou Anthony Gavalas Robin Lovell-Badge Eumorphia Remboutsika |
| author_sort | Maria Poulou |
| collection | DOAJ |
| description | <h4>Background</h4>Present technology uses mostly chimeric proteins as regulators and hormones or antibiotics as signals to induce spatial and temporal gene expression.<h4>Methodology/principal findings</h4>Here, we show that a chromosomally integrated yeast 'Leu3p-alpha-IotaRhoMu' system constitutes a ligand-inducible regulatory "off-on" genetic switch with an extensively dynamic action area. We find that Leu3p acts as an active transcriptional repressor in the absence and as an activator in the presence of alpha-isopropylmalate (alpha-IotaRhoMu) in primary fibroblasts isolated from double transgenic mouse embryos bearing ubiquitously expressing Leu3p and a Leu3p regulated GFP reporter. In the absence of the branched amino acid biosynthetic pathway in animals, metabolically stable alpha-IPM presents an EC(50) equal to 0.8837 mM and fast "OFF-ON" kinetics (t(50)ON = 43 min, t(50)OFF = 2.18 h), it enters the cells via passive diffusion, while it is non-toxic to mammalian cells and to fertilized mouse eggs cultured ex vivo.<h4>Conclusions/significance</h4>Our results demonstrate that the 'Leu3p-alpha-IotaRhoMu' constitutes a simpler and safer system for inducible gene expression in biomedical applications. |
| format | Article |
| id | doaj-art-03a0d5a5fbb94dde811dcb203e529d54 |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2010-08-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-03a0d5a5fbb94dde811dcb203e529d542025-08-20T02:01:58ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-08-0158e1248810.1371/journal.pone.0012488Development of a chromosomally integrated metabolite-inducible Leu3p-alpha-IPM "off-on" gene switch.Maria PoulouDonald BellKostas BozonelosMaria AlexiouAnthony GavalasRobin Lovell-BadgeEumorphia Remboutsika<h4>Background</h4>Present technology uses mostly chimeric proteins as regulators and hormones or antibiotics as signals to induce spatial and temporal gene expression.<h4>Methodology/principal findings</h4>Here, we show that a chromosomally integrated yeast 'Leu3p-alpha-IotaRhoMu' system constitutes a ligand-inducible regulatory "off-on" genetic switch with an extensively dynamic action area. We find that Leu3p acts as an active transcriptional repressor in the absence and as an activator in the presence of alpha-isopropylmalate (alpha-IotaRhoMu) in primary fibroblasts isolated from double transgenic mouse embryos bearing ubiquitously expressing Leu3p and a Leu3p regulated GFP reporter. In the absence of the branched amino acid biosynthetic pathway in animals, metabolically stable alpha-IPM presents an EC(50) equal to 0.8837 mM and fast "OFF-ON" kinetics (t(50)ON = 43 min, t(50)OFF = 2.18 h), it enters the cells via passive diffusion, while it is non-toxic to mammalian cells and to fertilized mouse eggs cultured ex vivo.<h4>Conclusions/significance</h4>Our results demonstrate that the 'Leu3p-alpha-IotaRhoMu' constitutes a simpler and safer system for inducible gene expression in biomedical applications.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0012488&type=printable |
| spellingShingle | Maria Poulou Donald Bell Kostas Bozonelos Maria Alexiou Anthony Gavalas Robin Lovell-Badge Eumorphia Remboutsika Development of a chromosomally integrated metabolite-inducible Leu3p-alpha-IPM "off-on" gene switch. PLoS ONE |
| title | Development of a chromosomally integrated metabolite-inducible Leu3p-alpha-IPM "off-on" gene switch. |
| title_full | Development of a chromosomally integrated metabolite-inducible Leu3p-alpha-IPM "off-on" gene switch. |
| title_fullStr | Development of a chromosomally integrated metabolite-inducible Leu3p-alpha-IPM "off-on" gene switch. |
| title_full_unstemmed | Development of a chromosomally integrated metabolite-inducible Leu3p-alpha-IPM "off-on" gene switch. |
| title_short | Development of a chromosomally integrated metabolite-inducible Leu3p-alpha-IPM "off-on" gene switch. |
| title_sort | development of a chromosomally integrated metabolite inducible leu3p alpha ipm off on gene switch |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0012488&type=printable |
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