PUBLICATION
Next-generation plasmids for transgenesis in zebrafish and beyond
- Authors
- Kemmler, C.L., Moran, H.R., Murray, B.F., Scoresby, A., Klem, J.R., Eckert, R.L., Lepovsky, E., Bertho, S., Nieuwenhuize, S., Burger, S., D'Agati, G., Betz, C., Puller, A.C., Felker, A., Ditrychova, K., Bötschi, S., Affolter, M., Rohner, N., Lovely, C.B., Kwan, K.M., Burger, A., Mosimann, C.
- ID
- ZDB-PUB-230329-46
- Date
- 2023
- Source
- Development (Cambridge, England) 150(8): (Journal)
- Registered Authors
- Affolter, Markus, Burger, Alexa, D'Agati, Gianluca, Kemmler, Cassie L., Kwan, Kristen, Lovely, Ben, Mosimann, Christian, Nieuwenhuize, Susan, Rohner, Nicolas
- Keywords
- Cavefish, Enhancer Discovery, Flurophores, Trangenesis Marker, Transgenesis, Zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- DNA Transposable Elements/genetics
- Gene Transfer Techniques*
- Mice
- Plasmids/genetics
- Promoter Regions, Genetic/genetics
- Zebrafish*/genetics
- Zebrafish*/metabolism
- PubMed
- 36975217 Full text @ Development
Citation
Kemmler, C.L., Moran, H.R., Murray, B.F., Scoresby, A., Klem, J.R., Eckert, R.L., Lepovsky, E., Bertho, S., Nieuwenhuize, S., Burger, S., D'Agati, G., Betz, C., Puller, A.C., Felker, A., Ditrychova, K., Bötschi, S., Affolter, M., Rohner, N., Lovely, C.B., Kwan, K.M., Burger, A., Mosimann, C. (2023) Next-generation plasmids for transgenesis in zebrafish and beyond. Development (Cambridge, England). 150(8):.
Abstract
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 beta-globin 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 mCerulean 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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping