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Gene-specific cell labeling using MiMIC transposons

Binary expression systems such as GAL4/UAS, LexA/LexAop and QF/QUAS have greatly enhanced the power of Drosophila as a model organism by allowing spatio-temporal manipulation of gene function as well as cell and neural circuit function. Tissue-specific expression of these heterologous transcription...

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Autores principales: Gnerer, Joshua P., Venken, Koen J. T., Dierick, Herman A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4417149/
https://www.ncbi.nlm.nih.gov/pubmed/25712101
http://dx.doi.org/10.1093/nar/gkv113
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author Gnerer, Joshua P.
Venken, Koen J. T.
Dierick, Herman A.
author_facet Gnerer, Joshua P.
Venken, Koen J. T.
Dierick, Herman A.
author_sort Gnerer, Joshua P.
collection PubMed
description Binary expression systems such as GAL4/UAS, LexA/LexAop and QF/QUAS have greatly enhanced the power of Drosophila as a model organism by allowing spatio-temporal manipulation of gene function as well as cell and neural circuit function. Tissue-specific expression of these heterologous transcription factors relies on random transposon integration near enhancers or promoters that drive the binary transcription factor embedded in the transposon. Alternatively, gene-specific promoter elements are directly fused to the binary factor within the transposon followed by random or site-specific integration. However, such insertions do not consistently recapitulate endogenous expression. We used Minos-Mediated Integration Cassette (MiMIC) transposons to convert host loci into reliable gene-specific binary effectors. MiMIC transposons allow recombinase-mediated cassette exchange to modify the transposon content. We developed novel exchange cassettes to convert coding intronic MiMIC insertions into gene-specific binary factor protein-traps. In addition, we expanded the set of binary factor exchange cassettes available for non-coding intronic MiMIC insertions. We show that binary factor conversions of different insertions in the same locus have indistinguishable expression patterns, suggesting that they reliably reflect endogenous gene expression. We show the efficacy and broad applicability of these new tools by dissecting the cellular expression patterns of the Drosophila serotonin receptor gene family.
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spelling pubmed-44171492015-05-12 Gene-specific cell labeling using MiMIC transposons Gnerer, Joshua P. Venken, Koen J. T. Dierick, Herman A. Nucleic Acids Res Methods Online Binary expression systems such as GAL4/UAS, LexA/LexAop and QF/QUAS have greatly enhanced the power of Drosophila as a model organism by allowing spatio-temporal manipulation of gene function as well as cell and neural circuit function. Tissue-specific expression of these heterologous transcription factors relies on random transposon integration near enhancers or promoters that drive the binary transcription factor embedded in the transposon. Alternatively, gene-specific promoter elements are directly fused to the binary factor within the transposon followed by random or site-specific integration. However, such insertions do not consistently recapitulate endogenous expression. We used Minos-Mediated Integration Cassette (MiMIC) transposons to convert host loci into reliable gene-specific binary effectors. MiMIC transposons allow recombinase-mediated cassette exchange to modify the transposon content. We developed novel exchange cassettes to convert coding intronic MiMIC insertions into gene-specific binary factor protein-traps. In addition, we expanded the set of binary factor exchange cassettes available for non-coding intronic MiMIC insertions. We show that binary factor conversions of different insertions in the same locus have indistinguishable expression patterns, suggesting that they reliably reflect endogenous gene expression. We show the efficacy and broad applicability of these new tools by dissecting the cellular expression patterns of the Drosophila serotonin receptor gene family. Oxford University Press 2015-04-30 2015-02-20 /pmc/articles/PMC4417149/ /pubmed/25712101 http://dx.doi.org/10.1093/nar/gkv113 Text en © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Gnerer, Joshua P.
Venken, Koen J. T.
Dierick, Herman A.
Gene-specific cell labeling using MiMIC transposons
title Gene-specific cell labeling using MiMIC transposons
title_full Gene-specific cell labeling using MiMIC transposons
title_fullStr Gene-specific cell labeling using MiMIC transposons
title_full_unstemmed Gene-specific cell labeling using MiMIC transposons
title_short Gene-specific cell labeling using MiMIC transposons
title_sort gene-specific cell labeling using mimic transposons
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4417149/
https://www.ncbi.nlm.nih.gov/pubmed/25712101
http://dx.doi.org/10.1093/nar/gkv113
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