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Engineering Photoactivatability in Genetically Encoded Voltage and pH Indicators
Genetically-encoded indicators of neuronal activity enable the labeling of a genetically defined population of neurons to optically monitor their activities. However, researchers often find difficulties in identifying relevant signals from excessive background fluorescence. A photoactivatable versio...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6828978/ https://www.ncbi.nlm.nih.gov/pubmed/31736711 http://dx.doi.org/10.3389/fncel.2019.00482 |
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author | Lee, Sungmoo Song, Yoon-Kyu Baker, Bradley J. |
author_facet | Lee, Sungmoo Song, Yoon-Kyu Baker, Bradley J. |
author_sort | Lee, Sungmoo |
collection | PubMed |
description | Genetically-encoded indicators of neuronal activity enable the labeling of a genetically defined population of neurons to optically monitor their activities. However, researchers often find difficulties in identifying relevant signals from excessive background fluorescence. A photoactivatable version of a genetically encoded calcium indicator, sPA-GCaMP6f is a good example of circumventing such an obstacle by limiting the fluorescence to a region of interest defined by the user. Here, we apply this strategy to genetically encoded voltage (GEVI) and pH (GEPI) indicators. Three photoactivatable GEVI candidates were considered. The first one used a circularly-permuted fluorescent protein, the second design involved a Förster resonance energy transfer (FRET) pair, and the third approach employed a pH-sensitive variant of GFP, ecliptic pHluorin. The candidate with a variant of ecliptic pHluorin exhibited photoactivation and a voltage-dependent fluorescence change. This effort also yielded a pH-sensitive photoactivatable GFP that varies its brightness in response to intracellular pH changes. |
format | Online Article Text |
id | pubmed-6828978 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-68289782019-11-15 Engineering Photoactivatability in Genetically Encoded Voltage and pH Indicators Lee, Sungmoo Song, Yoon-Kyu Baker, Bradley J. Front Cell Neurosci Cellular Neuroscience Genetically-encoded indicators of neuronal activity enable the labeling of a genetically defined population of neurons to optically monitor their activities. However, researchers often find difficulties in identifying relevant signals from excessive background fluorescence. A photoactivatable version of a genetically encoded calcium indicator, sPA-GCaMP6f is a good example of circumventing such an obstacle by limiting the fluorescence to a region of interest defined by the user. Here, we apply this strategy to genetically encoded voltage (GEVI) and pH (GEPI) indicators. Three photoactivatable GEVI candidates were considered. The first one used a circularly-permuted fluorescent protein, the second design involved a Förster resonance energy transfer (FRET) pair, and the third approach employed a pH-sensitive variant of GFP, ecliptic pHluorin. The candidate with a variant of ecliptic pHluorin exhibited photoactivation and a voltage-dependent fluorescence change. This effort also yielded a pH-sensitive photoactivatable GFP that varies its brightness in response to intracellular pH changes. Frontiers Media S.A. 2019-10-29 /pmc/articles/PMC6828978/ /pubmed/31736711 http://dx.doi.org/10.3389/fncel.2019.00482 Text en Copyright © 2019 Lee, Song and Baker. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Cellular Neuroscience Lee, Sungmoo Song, Yoon-Kyu Baker, Bradley J. Engineering Photoactivatability in Genetically Encoded Voltage and pH Indicators |
title | Engineering Photoactivatability in Genetically Encoded Voltage and pH Indicators |
title_full | Engineering Photoactivatability in Genetically Encoded Voltage and pH Indicators |
title_fullStr | Engineering Photoactivatability in Genetically Encoded Voltage and pH Indicators |
title_full_unstemmed | Engineering Photoactivatability in Genetically Encoded Voltage and pH Indicators |
title_short | Engineering Photoactivatability in Genetically Encoded Voltage and pH Indicators |
title_sort | engineering photoactivatability in genetically encoded voltage and ph indicators |
topic | Cellular Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6828978/ https://www.ncbi.nlm.nih.gov/pubmed/31736711 http://dx.doi.org/10.3389/fncel.2019.00482 |
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