Next-generation plasmids for transgenesis in zebrafish and beyond

Kemmler Cassie L: Moran, MoHannah R. and F. Murray, Brooke and Scoresby, Aaron and Klem, John R. and Eckert, Rachel L. and Lepovsky, Elizabeth and Bertho, Sylvain and Nieuwenhuize, Susan and Burger, Sibylle and D'Agati, Gianluca and Betz, Charles and Puller, Ann-Christin and Felker, Anastasia and Ditrychová, Karolina and Bötschi, Seraina and Affolter, Markus and Rohner, Nicolas and Lovely, C. Ben and Kwan M, Kristen and Burger, Alexa and Mosimann, Christian. (2022) Next-generation plasmids for transgenesis in zebrafish and beyond.

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Official URL: https://edoc.unibas.ch/91779/

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Transgenesis is an essential technique for any genetic model. Tol2-based transgenesis paired with Gateway-compatible vector collections has transformed zebrafish transgenesis with an accessible, modular system. Here, we established several next-generation transgenesis tools for zebrafish and other species to expand and enhance transgenic applications. To facilitate gene-regulatory element testing, we generated Gateway middle entry vectors harboring the small mouse betaglobin minimal promoter coupled to several fluorophores, CreERT2, and Gal4. To extend the color spectrum for transgenic applications, we established middle entry vectors encoding the bright, blue-fluorescent protein Cerulean and mApple as an alternative red fluorophore. We present a series of p2A peptide-based 3' vectors with different fluorophores and subcellular localizations to co-label cells expressing proteins of interest. Lastly, we established Tol2 destination vectors carrying the zebrafish exorh promoter driving different fluorophores as a pineal gland-specific transgenesis marker active prior to hatching and through adulthood. exorh-based reporters and transgenesis markers also drive specific pineal gland expression in the eye-less cavefish (Astyanax). Together, our vectors provide versatile reagents for transgenesis applications in zebrafish, cavefish, and other models.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Growth & Development > Cell Biology (Affolter)
UniBasel Contributors:Affolter, Markus
Item Type:Preprint
Note:Publication type according to Uni Basel Research Database: Discussion paper / Internet publication
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Last Modified:21 Dec 2022 08:30
Deposited On:21 Dec 2022 08:30

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