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The inducible lac operator-repressor system is functional in zebrafish cells

View ORCID ProfileSierra S. Nishizaki, View ORCID ProfileTorrin L. McDonald, View ORCID ProfileGregory A. Farnum, View ORCID ProfileMonica J. Holmes, View ORCID ProfileMelissa L. Drexel, View ORCID ProfileJessica A. Switzenberg, View ORCID ProfileAlan P. Boyle
doi: https://doi.org/10.1101/2020.02.10.939223
Sierra S. Nishizaki
1Department of Human Genetics, University of Michigan, Ann Arbor, MI, USA 48109
2Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI, USA 48109
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  • ORCID record for Sierra S. Nishizaki
Torrin L. McDonald
1Department of Human Genetics, University of Michigan, Ann Arbor, MI, USA 48109
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Gregory A. Farnum
2Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI, USA 48109
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Monica J. Holmes
2Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI, USA 48109
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  • ORCID record for Monica J. Holmes
Melissa L. Drexel
1Department of Human Genetics, University of Michigan, Ann Arbor, MI, USA 48109
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  • ORCID record for Melissa L. Drexel
Jessica A. Switzenberg
2Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI, USA 48109
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  • ORCID record for Jessica A. Switzenberg
Alan P. Boyle
1Department of Human Genetics, University of Michigan, Ann Arbor, MI, USA 48109
2Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI, USA 48109
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  • For correspondence: apboyle@umich.edu
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Abstract

Zebrafish have been 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, a component of the E. Coli lac operon, has been adapted for use in many other species to study the inducible modulation of gene expression, but has not previously been shown to function in zebrafish. 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. Additionally we show the first evidence that the cytomegalovirus (CMV) enhancer is active in zebrafish cells and enhances eGFP expression ubiquitously in zebrafish embryos. Our results lay the groundwork for the development of lac-based reporter assays in zebrafish, and add to the tools available for investigating dynamic gene expression in embryogenesis. We believe that 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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Posted February 10, 2020.
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The inducible lac operator-repressor system is functional in zebrafish cells
Sierra S. Nishizaki, Torrin L. McDonald, Gregory A. Farnum, Monica J. Holmes, Melissa L. Drexel, Jessica A. Switzenberg, Alan P. Boyle
bioRxiv 2020.02.10.939223; doi: https://doi.org/10.1101/2020.02.10.939223
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The inducible lac operator-repressor system is functional in zebrafish cells
Sierra S. Nishizaki, Torrin L. McDonald, Gregory A. Farnum, Monica J. Holmes, Melissa L. Drexel, Jessica A. Switzenberg, Alan P. Boyle
bioRxiv 2020.02.10.939223; doi: https://doi.org/10.1101/2020.02.10.939223

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