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An optogenetic gene expression system with rapid activation and deactivation kinetics

Optogenetic gene expression systems can control transcription with spatial and temporal detail unequaled with traditional inducible promoter systems. However, current eukaryotic light-gated transcription systems are limited by toxicity, dynamic range, or slow activation/deactivation. Here we present...

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Detalles Bibliográficos
Autores principales: Motta-Mena, Laura B., Reade, Anna, Mallory, Michael J., Glantz, Spencer, Weiner, Orion D., Lynch, Kristen W., Gardner, Kevin H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3944926/
https://www.ncbi.nlm.nih.gov/pubmed/24413462
http://dx.doi.org/10.1038/nchembio.1430
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author Motta-Mena, Laura B.
Reade, Anna
Mallory, Michael J.
Glantz, Spencer
Weiner, Orion D.
Lynch, Kristen W.
Gardner, Kevin H.
author_facet Motta-Mena, Laura B.
Reade, Anna
Mallory, Michael J.
Glantz, Spencer
Weiner, Orion D.
Lynch, Kristen W.
Gardner, Kevin H.
author_sort Motta-Mena, Laura B.
collection PubMed
description Optogenetic gene expression systems can control transcription with spatial and temporal detail unequaled with traditional inducible promoter systems. However, current eukaryotic light-gated transcription systems are limited by toxicity, dynamic range, or slow activation/deactivation. Here we present an optogenetic gene expression system that addresses these shortcomings and demonstrate its broad utility. Our approach utilizes an engineered version of EL222, a bacterial Light-Oxygen-Voltage (LOV) protein that binds DNA when illuminated with blue light. The system has a large (>100-fold) dynamic range of protein expression, rapid activation (< 10 s) and deactivation kinetics (< 50 s), and a highly linear response to light. With this system, we achieve light-gated transcription in several mammalian cell lines and intact zebrafish embryos with minimal basal gene activation and toxicity. Our approach provides a powerful new tool for optogenetic control of gene expression in space and time.
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spelling pubmed-39449262014-09-01 An optogenetic gene expression system with rapid activation and deactivation kinetics Motta-Mena, Laura B. Reade, Anna Mallory, Michael J. Glantz, Spencer Weiner, Orion D. Lynch, Kristen W. Gardner, Kevin H. Nat Chem Biol Article Optogenetic gene expression systems can control transcription with spatial and temporal detail unequaled with traditional inducible promoter systems. However, current eukaryotic light-gated transcription systems are limited by toxicity, dynamic range, or slow activation/deactivation. Here we present an optogenetic gene expression system that addresses these shortcomings and demonstrate its broad utility. Our approach utilizes an engineered version of EL222, a bacterial Light-Oxygen-Voltage (LOV) protein that binds DNA when illuminated with blue light. The system has a large (>100-fold) dynamic range of protein expression, rapid activation (< 10 s) and deactivation kinetics (< 50 s), and a highly linear response to light. With this system, we achieve light-gated transcription in several mammalian cell lines and intact zebrafish embryos with minimal basal gene activation and toxicity. Our approach provides a powerful new tool for optogenetic control of gene expression in space and time. 2014-01-12 2014-03 /pmc/articles/PMC3944926/ /pubmed/24413462 http://dx.doi.org/10.1038/nchembio.1430 Text en Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Motta-Mena, Laura B.
Reade, Anna
Mallory, Michael J.
Glantz, Spencer
Weiner, Orion D.
Lynch, Kristen W.
Gardner, Kevin H.
An optogenetic gene expression system with rapid activation and deactivation kinetics
title An optogenetic gene expression system with rapid activation and deactivation kinetics
title_full An optogenetic gene expression system with rapid activation and deactivation kinetics
title_fullStr An optogenetic gene expression system with rapid activation and deactivation kinetics
title_full_unstemmed An optogenetic gene expression system with rapid activation and deactivation kinetics
title_short An optogenetic gene expression system with rapid activation and deactivation kinetics
title_sort optogenetic gene expression system with rapid activation and deactivation kinetics
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3944926/
https://www.ncbi.nlm.nih.gov/pubmed/24413462
http://dx.doi.org/10.1038/nchembio.1430
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