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Spatial and temporal control of expression with light-gated LOV-LexA

The ability to drive expression of exogenous genes in different tissues and cell types, under the control of specific enhancers, has been crucial for discovery in biology. While many enhancers drive expression broadly, several genetic tools were developed to obtain access to isolated cell types. Stu...

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Autores principales: Ribeiro, Inês M A, Eßbauer, Wolfgang, Kutlesa, Romina, Borst, Alexander
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9526042/
https://www.ncbi.nlm.nih.gov/pubmed/35876796
http://dx.doi.org/10.1093/g3journal/jkac178
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author Ribeiro, Inês M A
Eßbauer, Wolfgang
Kutlesa, Romina
Borst, Alexander
author_facet Ribeiro, Inês M A
Eßbauer, Wolfgang
Kutlesa, Romina
Borst, Alexander
author_sort Ribeiro, Inês M A
collection PubMed
description The ability to drive expression of exogenous genes in different tissues and cell types, under the control of specific enhancers, has been crucial for discovery in biology. While many enhancers drive expression broadly, several genetic tools were developed to obtain access to isolated cell types. Studies of spatially organized neuropiles in the central nervous system of fruit flies have raised the need for a system that targets subsets of cells within a single neuronal type, a feat currently dependent on stochastic flip-out methods. To access the same cells within a given expression pattern consistently across fruit flies, we developed the light-gated expression system LOV-LexA. We combined the bacterial LexA transcription factor with the plant-derived light, oxygen, or voltage photosensitive domain and a fluorescent protein. Exposure to blue light uncages a nuclear localizing signal in the C-terminal of the light, oxygen, or voltage domain and leads to the translocation of LOV-LexA to the nucleus, with the subsequent initiation of transcription. LOV-LexA enables spatial and temporal control of expression of transgenes under LexAop sequences in larval fat body and pupal and adult neurons with blue light. The LOV-LexA tool is ready to use with GAL4 and Split-GAL4 drivers in its current form and constitutes another layer of intersectional genetics that provides light-controlled genetic access to specific cells across flies.
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spelling pubmed-95260422022-10-03 Spatial and temporal control of expression with light-gated LOV-LexA Ribeiro, Inês M A Eßbauer, Wolfgang Kutlesa, Romina Borst, Alexander G3 (Bethesda) Investigation The ability to drive expression of exogenous genes in different tissues and cell types, under the control of specific enhancers, has been crucial for discovery in biology. While many enhancers drive expression broadly, several genetic tools were developed to obtain access to isolated cell types. Studies of spatially organized neuropiles in the central nervous system of fruit flies have raised the need for a system that targets subsets of cells within a single neuronal type, a feat currently dependent on stochastic flip-out methods. To access the same cells within a given expression pattern consistently across fruit flies, we developed the light-gated expression system LOV-LexA. We combined the bacterial LexA transcription factor with the plant-derived light, oxygen, or voltage photosensitive domain and a fluorescent protein. Exposure to blue light uncages a nuclear localizing signal in the C-terminal of the light, oxygen, or voltage domain and leads to the translocation of LOV-LexA to the nucleus, with the subsequent initiation of transcription. LOV-LexA enables spatial and temporal control of expression of transgenes under LexAop sequences in larval fat body and pupal and adult neurons with blue light. The LOV-LexA tool is ready to use with GAL4 and Split-GAL4 drivers in its current form and constitutes another layer of intersectional genetics that provides light-controlled genetic access to specific cells across flies. Oxford University Press 2022-07-25 /pmc/articles/PMC9526042/ /pubmed/35876796 http://dx.doi.org/10.1093/g3journal/jkac178 Text en © The Author(s) 2022. Published by Oxford University Press on behalf of Genetics Society of America. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Investigation
Ribeiro, Inês M A
Eßbauer, Wolfgang
Kutlesa, Romina
Borst, Alexander
Spatial and temporal control of expression with light-gated LOV-LexA
title Spatial and temporal control of expression with light-gated LOV-LexA
title_full Spatial and temporal control of expression with light-gated LOV-LexA
title_fullStr Spatial and temporal control of expression with light-gated LOV-LexA
title_full_unstemmed Spatial and temporal control of expression with light-gated LOV-LexA
title_short Spatial and temporal control of expression with light-gated LOV-LexA
title_sort spatial and temporal control of expression with light-gated lov-lexa
topic Investigation
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9526042/
https://www.ncbi.nlm.nih.gov/pubmed/35876796
http://dx.doi.org/10.1093/g3journal/jkac178
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