pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH.
We report the development of a genetically encodable and ratiometic pH probe named "pHlash" that utilizes Bioluminescence Resonance Energy Transfer (BRET) rather than fluorescence excitation. The pHlash sensor-composed of a donor luciferase that is genetically fused to a Venus fluorophore-...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22905204/?tool=EBI |
| _version_ | 1818573838326169600 |
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| author | Yunfei Zhang Qiguang Xie J Brian Robertson Carl Hirschie Johnson |
| author_facet | Yunfei Zhang Qiguang Xie J Brian Robertson Carl Hirschie Johnson |
| author_sort | Yunfei Zhang |
| collection | DOAJ |
| description | We report the development of a genetically encodable and ratiometic pH probe named "pHlash" that utilizes Bioluminescence Resonance Energy Transfer (BRET) rather than fluorescence excitation. The pHlash sensor-composed of a donor luciferase that is genetically fused to a Venus fluorophore-exhibits pH dependence of its spectral emission in vitro. When expressed in either yeast or mammalian cells, pHlash reports basal pH and cytosolic acidification in vivo. Its spectral ratio response is H(+) specific; neither Ca(++), Mg(++), Na(+), nor K(+) changes the spectral form of its luminescence emission. Moreover, it can be used to image pH in single cells. This is the first BRET-based sensor of H(+) ions, and it should allow the approximation of pH in cytosolic and organellar compartments in applications where current pH probes are inadequate. |
| first_indexed | 2024-12-15T00:18:15Z |
| format | Article |
| id | doaj.art-3f36e15c5aa94d69956ca02ceee93b69 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-15T00:18:15Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-3f36e15c5aa94d69956ca02ceee93b692022-12-21T22:42:24ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0178e4307210.1371/journal.pone.0043072pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH.Yunfei ZhangQiguang XieJ Brian RobertsonCarl Hirschie JohnsonWe report the development of a genetically encodable and ratiometic pH probe named "pHlash" that utilizes Bioluminescence Resonance Energy Transfer (BRET) rather than fluorescence excitation. The pHlash sensor-composed of a donor luciferase that is genetically fused to a Venus fluorophore-exhibits pH dependence of its spectral emission in vitro. When expressed in either yeast or mammalian cells, pHlash reports basal pH and cytosolic acidification in vivo. Its spectral ratio response is H(+) specific; neither Ca(++), Mg(++), Na(+), nor K(+) changes the spectral form of its luminescence emission. Moreover, it can be used to image pH in single cells. This is the first BRET-based sensor of H(+) ions, and it should allow the approximation of pH in cytosolic and organellar compartments in applications where current pH probes are inadequate.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22905204/?tool=EBI |
| spellingShingle | Yunfei Zhang Qiguang Xie J Brian Robertson Carl Hirschie Johnson pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH. PLoS ONE |
| title | pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH. |
| title_full | pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH. |
| title_fullStr | pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH. |
| title_full_unstemmed | pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH. |
| title_short | pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH. |
| title_sort | phlash a new genetically encoded and ratiometric luminescence sensor of intracellular ph |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22905204/?tool=EBI |
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