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KappaBle fluorescent reporter mice enable low-background single-cell detection of NF-κB transcriptional activity in vivo
Nuclear factor-κB (NF-κB) is a transcription factor with a key role in a great variety of cellular processes from embryonic development to immunity, the outcome of which depends on the fine-tuning of NF-κB activity. The development of sensitive and faithful reporter systems to accurately monitor the...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group US
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9259492/ https://www.ncbi.nlm.nih.gov/pubmed/35589985 http://dx.doi.org/10.1038/s41385-022-00525-8 |
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author | Tortola, Luigi Piattini, Federica Hausmann, Annika Ampenberger, Franziska Rosenwald, Esther Heer, Sebastian Hardt, Wolf-Dietrich Rülicke, Thomas Kisielow, Jan Kopf, Manfred |
author_facet | Tortola, Luigi Piattini, Federica Hausmann, Annika Ampenberger, Franziska Rosenwald, Esther Heer, Sebastian Hardt, Wolf-Dietrich Rülicke, Thomas Kisielow, Jan Kopf, Manfred |
author_sort | Tortola, Luigi |
collection | PubMed |
description | Nuclear factor-κB (NF-κB) is a transcription factor with a key role in a great variety of cellular processes from embryonic development to immunity, the outcome of which depends on the fine-tuning of NF-κB activity. The development of sensitive and faithful reporter systems to accurately monitor the activation status of this transcription factor is therefore desirable. To address this need, over the years a number of different approaches have been used to generate NF-κB reporter mice, which can be broadly subdivided into bioluminescence- and fluorescence-based systems. While the former enables whole-body visualization of the activation status of NF-κB, the latter have the potential to allow the analysis of NF-κB activity at single-cell level. However, fluorescence-based reporters frequently show poor sensitivity and excessive background or are incompatible with high-throughput flow cytometric analysis. In this work we describe the generation and analysis of ROSA26 knock-in NF-κB reporter (KappaBle) mice containing a destabilized EGFP, which showed sensitive, dynamic, and faithful monitoring of NF-κB transcriptional activity at the single-cell level of various cell types during inflammatory and infectious diseases. |
format | Online Article Text |
id | pubmed-9259492 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group US |
record_format | MEDLINE/PubMed |
spelling | pubmed-92594922022-07-08 KappaBle fluorescent reporter mice enable low-background single-cell detection of NF-κB transcriptional activity in vivo Tortola, Luigi Piattini, Federica Hausmann, Annika Ampenberger, Franziska Rosenwald, Esther Heer, Sebastian Hardt, Wolf-Dietrich Rülicke, Thomas Kisielow, Jan Kopf, Manfred Mucosal Immunol Article Nuclear factor-κB (NF-κB) is a transcription factor with a key role in a great variety of cellular processes from embryonic development to immunity, the outcome of which depends on the fine-tuning of NF-κB activity. The development of sensitive and faithful reporter systems to accurately monitor the activation status of this transcription factor is therefore desirable. To address this need, over the years a number of different approaches have been used to generate NF-κB reporter mice, which can be broadly subdivided into bioluminescence- and fluorescence-based systems. While the former enables whole-body visualization of the activation status of NF-κB, the latter have the potential to allow the analysis of NF-κB activity at single-cell level. However, fluorescence-based reporters frequently show poor sensitivity and excessive background or are incompatible with high-throughput flow cytometric analysis. In this work we describe the generation and analysis of ROSA26 knock-in NF-κB reporter (KappaBle) mice containing a destabilized EGFP, which showed sensitive, dynamic, and faithful monitoring of NF-κB transcriptional activity at the single-cell level of various cell types during inflammatory and infectious diseases. Nature Publishing Group US 2022-05-19 2022 /pmc/articles/PMC9259492/ /pubmed/35589985 http://dx.doi.org/10.1038/s41385-022-00525-8 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Tortola, Luigi Piattini, Federica Hausmann, Annika Ampenberger, Franziska Rosenwald, Esther Heer, Sebastian Hardt, Wolf-Dietrich Rülicke, Thomas Kisielow, Jan Kopf, Manfred KappaBle fluorescent reporter mice enable low-background single-cell detection of NF-κB transcriptional activity in vivo |
title | KappaBle fluorescent reporter mice enable low-background single-cell detection of NF-κB transcriptional activity in vivo |
title_full | KappaBle fluorescent reporter mice enable low-background single-cell detection of NF-κB transcriptional activity in vivo |
title_fullStr | KappaBle fluorescent reporter mice enable low-background single-cell detection of NF-κB transcriptional activity in vivo |
title_full_unstemmed | KappaBle fluorescent reporter mice enable low-background single-cell detection of NF-κB transcriptional activity in vivo |
title_short | KappaBle fluorescent reporter mice enable low-background single-cell detection of NF-κB transcriptional activity in vivo |
title_sort | kappable fluorescent reporter mice enable low-background single-cell detection of nf-κb transcriptional activity in vivo |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9259492/ https://www.ncbi.nlm.nih.gov/pubmed/35589985 http://dx.doi.org/10.1038/s41385-022-00525-8 |
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