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Generation of a Novel Nkx6-1 Venus Fusion Reporter Mouse Line

Nkx6-1 is a member of the Nkx family of homeodomain transcription factors (TFs) that regulates motor neuron development, neuron specification and pancreatic endocrine and β-cell differentiation. To facilitate the isolation and tracking of Nkx6-1-expressing cells, we have generated a novel Nkx6-1 Ven...

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Autores principales: Burtscher, Ingo, Tarquis-Medina, Marta, Salinno, Ciro, Schirge, Silvia, Beckenbauer, Julia, Bakhti, Mostafa, Lickert, Heiko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036392/
https://www.ncbi.nlm.nih.gov/pubmed/33810480
http://dx.doi.org/10.3390/ijms22073434
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author Burtscher, Ingo
Tarquis-Medina, Marta
Salinno, Ciro
Schirge, Silvia
Beckenbauer, Julia
Bakhti, Mostafa
Lickert, Heiko
author_facet Burtscher, Ingo
Tarquis-Medina, Marta
Salinno, Ciro
Schirge, Silvia
Beckenbauer, Julia
Bakhti, Mostafa
Lickert, Heiko
author_sort Burtscher, Ingo
collection PubMed
description Nkx6-1 is a member of the Nkx family of homeodomain transcription factors (TFs) that regulates motor neuron development, neuron specification and pancreatic endocrine and β-cell differentiation. To facilitate the isolation and tracking of Nkx6-1-expressing cells, we have generated a novel Nkx6-1 Venus fusion (Nkx6-1-VF) reporter allele. The Nkx6-1-VF knock-in reporter is regulated by endogenous cis-regulatory elements of Nkx6-1 and the fluorescent protein fusion does not interfere with the TF function, as homozygous mice are viable and fertile. The nuclear localization of Nkx6-1-VF protein reflects the endogenous Nkx6-1 protein distribution. During embryonic pancreas development, the reporter protein marks the pancreatic ductal progenitors and the endocrine lineage, but is absent in the exocrine compartment. As expected, the levels of Nkx6-1-VF reporter are upregulated upon β-cell differentiation during the major wave of endocrinogenesis. In the adult islets of Langerhans, the reporter protein is exclusively found in insulin-secreting β-cells. Importantly, the Venus reporter activities allow successful tracking of β-cells in live-cell imaging and their specific isolation by flow sorting. In summary, the generation of the Nkx6-1-VF reporter line reflects the expression pattern and dynamics of the endogenous protein and thus provides a unique tool to study the spatio-temporal expression pattern of this TF during organ development and enables isolation and tracking of Nkx6-1-expressing cells such as pancreatic β-cells, but also neurons and motor neurons in health and disease.
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spelling pubmed-80363922021-04-12 Generation of a Novel Nkx6-1 Venus Fusion Reporter Mouse Line Burtscher, Ingo Tarquis-Medina, Marta Salinno, Ciro Schirge, Silvia Beckenbauer, Julia Bakhti, Mostafa Lickert, Heiko Int J Mol Sci Article Nkx6-1 is a member of the Nkx family of homeodomain transcription factors (TFs) that regulates motor neuron development, neuron specification and pancreatic endocrine and β-cell differentiation. To facilitate the isolation and tracking of Nkx6-1-expressing cells, we have generated a novel Nkx6-1 Venus fusion (Nkx6-1-VF) reporter allele. The Nkx6-1-VF knock-in reporter is regulated by endogenous cis-regulatory elements of Nkx6-1 and the fluorescent protein fusion does not interfere with the TF function, as homozygous mice are viable and fertile. The nuclear localization of Nkx6-1-VF protein reflects the endogenous Nkx6-1 protein distribution. During embryonic pancreas development, the reporter protein marks the pancreatic ductal progenitors and the endocrine lineage, but is absent in the exocrine compartment. As expected, the levels of Nkx6-1-VF reporter are upregulated upon β-cell differentiation during the major wave of endocrinogenesis. In the adult islets of Langerhans, the reporter protein is exclusively found in insulin-secreting β-cells. Importantly, the Venus reporter activities allow successful tracking of β-cells in live-cell imaging and their specific isolation by flow sorting. In summary, the generation of the Nkx6-1-VF reporter line reflects the expression pattern and dynamics of the endogenous protein and thus provides a unique tool to study the spatio-temporal expression pattern of this TF during organ development and enables isolation and tracking of Nkx6-1-expressing cells such as pancreatic β-cells, but also neurons and motor neurons in health and disease. MDPI 2021-03-26 /pmc/articles/PMC8036392/ /pubmed/33810480 http://dx.doi.org/10.3390/ijms22073434 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Article
Burtscher, Ingo
Tarquis-Medina, Marta
Salinno, Ciro
Schirge, Silvia
Beckenbauer, Julia
Bakhti, Mostafa
Lickert, Heiko
Generation of a Novel Nkx6-1 Venus Fusion Reporter Mouse Line
title Generation of a Novel Nkx6-1 Venus Fusion Reporter Mouse Line
title_full Generation of a Novel Nkx6-1 Venus Fusion Reporter Mouse Line
title_fullStr Generation of a Novel Nkx6-1 Venus Fusion Reporter Mouse Line
title_full_unstemmed Generation of a Novel Nkx6-1 Venus Fusion Reporter Mouse Line
title_short Generation of a Novel Nkx6-1 Venus Fusion Reporter Mouse Line
title_sort generation of a novel nkx6-1 venus fusion reporter mouse line
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036392/
https://www.ncbi.nlm.nih.gov/pubmed/33810480
http://dx.doi.org/10.3390/ijms22073434
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