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Efficient tissue-type specific expression of target genes in a tetracycline-controlled manner from the ubiquitously active Eef1a1 locus

Using an efficient gene targeting approach, we developed a novel mouse line that expresses the tetracycline-controlled transactivator (tTA) from the constitutively active Eef1a1 locus in a Cre recombinase-inducible manner. The temporally and spatially controlled expression of the EF1-LSL-tTA knockin...

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Autores principales: Sakamoto, Kazuhito, Rädler, Patrick D., Wehde, Barbara L., Triplett, Aleata A., Shrestha, Hridaya, Ferraiuolo, Rosa-Maria, Amari, Foued, Coppola, Vincenzo, Klinakis, Apostolos, Efstratiadis, Argiris, Wagner, Kay-Uwe
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6959320/
https://www.ncbi.nlm.nih.gov/pubmed/31937792
http://dx.doi.org/10.1038/s41598-019-57052-z
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author Sakamoto, Kazuhito
Rädler, Patrick D.
Wehde, Barbara L.
Triplett, Aleata A.
Shrestha, Hridaya
Ferraiuolo, Rosa-Maria
Amari, Foued
Coppola, Vincenzo
Klinakis, Apostolos
Efstratiadis, Argiris
Wagner, Kay-Uwe
author_facet Sakamoto, Kazuhito
Rädler, Patrick D.
Wehde, Barbara L.
Triplett, Aleata A.
Shrestha, Hridaya
Ferraiuolo, Rosa-Maria
Amari, Foued
Coppola, Vincenzo
Klinakis, Apostolos
Efstratiadis, Argiris
Wagner, Kay-Uwe
author_sort Sakamoto, Kazuhito
collection PubMed
description Using an efficient gene targeting approach, we developed a novel mouse line that expresses the tetracycline-controlled transactivator (tTA) from the constitutively active Eef1a1 locus in a Cre recombinase-inducible manner. The temporally and spatially controlled expression of the EF1-LSL-tTA knockin and activation of tTA-driven responder transgenes was tested using four transgenic lines that express Cre under tissue-specific promoters of the pancreas, mammary gland and other secretory tissues, as well as an interferon-inducible promoter. In all models, the endogenous Eef1a1 promoter facilitated a cell-type-specific activation of target genes at high levels without exogenous enhancer elements. The applicability of the EF1-LSL-tTA strain for biological experiments was tested in two studies related to mammary gland development and tumorigenesis. First, we validated the crucial role of active STAT5 as a survival factor for functionally differentiated epithelial cells by expressing a hyperactive STAT5 mutant in the mammary gland during postlactational remodeling. In a second experiment, we assessed the ability of the EF1-tTA to initiate tumor formation through upregulation of mutant KRAS. The collective results show that the EF1-LSL-tTA knockin line is a versatile genetic tool that can be applied to constitutively express transgenes in specific cell types to examine their biological functions at defined developmental stages.
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spelling pubmed-69593202020-01-16 Efficient tissue-type specific expression of target genes in a tetracycline-controlled manner from the ubiquitously active Eef1a1 locus Sakamoto, Kazuhito Rädler, Patrick D. Wehde, Barbara L. Triplett, Aleata A. Shrestha, Hridaya Ferraiuolo, Rosa-Maria Amari, Foued Coppola, Vincenzo Klinakis, Apostolos Efstratiadis, Argiris Wagner, Kay-Uwe Sci Rep Article Using an efficient gene targeting approach, we developed a novel mouse line that expresses the tetracycline-controlled transactivator (tTA) from the constitutively active Eef1a1 locus in a Cre recombinase-inducible manner. The temporally and spatially controlled expression of the EF1-LSL-tTA knockin and activation of tTA-driven responder transgenes was tested using four transgenic lines that express Cre under tissue-specific promoters of the pancreas, mammary gland and other secretory tissues, as well as an interferon-inducible promoter. In all models, the endogenous Eef1a1 promoter facilitated a cell-type-specific activation of target genes at high levels without exogenous enhancer elements. The applicability of the EF1-LSL-tTA strain for biological experiments was tested in two studies related to mammary gland development and tumorigenesis. First, we validated the crucial role of active STAT5 as a survival factor for functionally differentiated epithelial cells by expressing a hyperactive STAT5 mutant in the mammary gland during postlactational remodeling. In a second experiment, we assessed the ability of the EF1-tTA to initiate tumor formation through upregulation of mutant KRAS. The collective results show that the EF1-LSL-tTA knockin line is a versatile genetic tool that can be applied to constitutively express transgenes in specific cell types to examine their biological functions at defined developmental stages. Nature Publishing Group UK 2020-01-14 /pmc/articles/PMC6959320/ /pubmed/31937792 http://dx.doi.org/10.1038/s41598-019-57052-z Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Sakamoto, Kazuhito
Rädler, Patrick D.
Wehde, Barbara L.
Triplett, Aleata A.
Shrestha, Hridaya
Ferraiuolo, Rosa-Maria
Amari, Foued
Coppola, Vincenzo
Klinakis, Apostolos
Efstratiadis, Argiris
Wagner, Kay-Uwe
Efficient tissue-type specific expression of target genes in a tetracycline-controlled manner from the ubiquitously active Eef1a1 locus
title Efficient tissue-type specific expression of target genes in a tetracycline-controlled manner from the ubiquitously active Eef1a1 locus
title_full Efficient tissue-type specific expression of target genes in a tetracycline-controlled manner from the ubiquitously active Eef1a1 locus
title_fullStr Efficient tissue-type specific expression of target genes in a tetracycline-controlled manner from the ubiquitously active Eef1a1 locus
title_full_unstemmed Efficient tissue-type specific expression of target genes in a tetracycline-controlled manner from the ubiquitously active Eef1a1 locus
title_short Efficient tissue-type specific expression of target genes in a tetracycline-controlled manner from the ubiquitously active Eef1a1 locus
title_sort efficient tissue-type specific expression of target genes in a tetracycline-controlled manner from the ubiquitously active eef1a1 locus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6959320/
https://www.ncbi.nlm.nih.gov/pubmed/31937792
http://dx.doi.org/10.1038/s41598-019-57052-z
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