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The Inducible lac Operator-Repressor System Is Functional in Zebrafish Cells

BACKGROUND: Zebrafish are a foundational model organism for studying the spatio-temporal activity of genes and their regulatory sequences. A variety of approaches are currently available for editing genes and modifying gene expression in zebrafish, including RNAi, Cre/lox, and CRISPR-Cas9. However,...

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Autores principales: Nishizaki, Sierra S., McDonald, Torrin L., Farnum, Gregory A., Holmes, Monica J., Drexel, Melissa L., Switzenberg, Jessica A., Boyle, Alan P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8249864/
https://www.ncbi.nlm.nih.gov/pubmed/34220959
http://dx.doi.org/10.3389/fgene.2021.683394
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author Nishizaki, Sierra S.
McDonald, Torrin L.
Farnum, Gregory A.
Holmes, Monica J.
Drexel, Melissa L.
Switzenberg, Jessica A.
Boyle, Alan P.
author_facet Nishizaki, Sierra S.
McDonald, Torrin L.
Farnum, Gregory A.
Holmes, Monica J.
Drexel, Melissa L.
Switzenberg, Jessica A.
Boyle, Alan P.
author_sort Nishizaki, Sierra S.
collection PubMed
description BACKGROUND: Zebrafish are a foundational model organism for studying the spatio-temporal activity of genes and their regulatory sequences. A variety of approaches are currently available for editing genes and modifying gene expression in zebrafish, including RNAi, Cre/lox, and CRISPR-Cas9. However, the lac operator-repressor system, an E. coli lac operon component which has been adapted for use in many other species and is a valuable, flexible tool for inducible modulation of gene expression studies, has not been previously tested in zebrafish. RESULTS: Here we demonstrate that the lac operator-repressor system robustly decreases expression of firefly luciferase in cultured zebrafish fibroblast cells. Our work establishes the lac operator-repressor system as a promising tool for the manipulation of gene expression in whole zebrafish. CONCLUSION: Our results lay the groundwork for the development of lac-based reporter assays in zebrafish, and adds to the tools available for investigating dynamic gene expression in embryogenesis. We believe this work will catalyze the development of new reporter assay systems to investigate uncharacterized regulatory elements and their cell-type specific activities.
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spelling pubmed-82498642021-07-03 The Inducible lac Operator-Repressor System Is Functional in Zebrafish Cells Nishizaki, Sierra S. McDonald, Torrin L. Farnum, Gregory A. Holmes, Monica J. Drexel, Melissa L. Switzenberg, Jessica A. Boyle, Alan P. Front Genet Genetics BACKGROUND: Zebrafish are a foundational model organism for studying the spatio-temporal activity of genes and their regulatory sequences. A variety of approaches are currently available for editing genes and modifying gene expression in zebrafish, including RNAi, Cre/lox, and CRISPR-Cas9. However, the lac operator-repressor system, an E. coli lac operon component which has been adapted for use in many other species and is a valuable, flexible tool for inducible modulation of gene expression studies, has not been previously tested in zebrafish. RESULTS: Here we demonstrate that the lac operator-repressor system robustly decreases expression of firefly luciferase in cultured zebrafish fibroblast cells. Our work establishes the lac operator-repressor system as a promising tool for the manipulation of gene expression in whole zebrafish. CONCLUSION: Our results lay the groundwork for the development of lac-based reporter assays in zebrafish, and adds to the tools available for investigating dynamic gene expression in embryogenesis. We believe this work will catalyze the development of new reporter assay systems to investigate uncharacterized regulatory elements and their cell-type specific activities. Frontiers Media S.A. 2021-06-18 /pmc/articles/PMC8249864/ /pubmed/34220959 http://dx.doi.org/10.3389/fgene.2021.683394 Text en Copyright © 2021 Nishizaki, McDonald, Farnum, Holmes, Drexel, Switzenberg and Boyle. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Genetics
Nishizaki, Sierra S.
McDonald, Torrin L.
Farnum, Gregory A.
Holmes, Monica J.
Drexel, Melissa L.
Switzenberg, Jessica A.
Boyle, Alan P.
The Inducible lac Operator-Repressor System Is Functional in Zebrafish Cells
title The Inducible lac Operator-Repressor System Is Functional in Zebrafish Cells
title_full The Inducible lac Operator-Repressor System Is Functional in Zebrafish Cells
title_fullStr The Inducible lac Operator-Repressor System Is Functional in Zebrafish Cells
title_full_unstemmed The Inducible lac Operator-Repressor System Is Functional in Zebrafish Cells
title_short The Inducible lac Operator-Repressor System Is Functional in Zebrafish Cells
title_sort inducible lac operator-repressor system is functional in zebrafish cells
topic Genetics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8249864/
https://www.ncbi.nlm.nih.gov/pubmed/34220959
http://dx.doi.org/10.3389/fgene.2021.683394
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