Next-generation plasmids for transgenesis in zebrafish and beyond
© 2023. Published by The Company of Biologists Ltd..
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 establish 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. Finally, we established Tol2 destination vectors carrying the zebrafish exorh promoter driving different fluorophores as a pineal gland-specific transgenesis marker that is active before 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.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:150 |
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| Enthalten in: |
Development (Cambridge, England) - 150(2023), 8 vom: 15. Apr. |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Kemmler, Cassie L [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 21.04.2023 Date Revised 04.02.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1242/dev.201531 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 245 | 1 | 0 | |a Next-generation plasmids for transgenesis in zebrafish and beyond |
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| 500 | |a Date Revised 04.02.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2023. Published by The Company of Biologists Ltd. | ||
| 520 | |a 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 establish 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. Finally, we established Tol2 destination vectors carrying the zebrafish exorh promoter driving different fluorophores as a pineal gland-specific transgenesis marker that is active before 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 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Research Support, U.S. Gov't, Non-P.H.S. | |
| 650 | 4 | |a Cavefish | |
| 650 | 4 | |a Enhancer discovery | |
| 650 | 4 | |a Flurophores | |
| 650 | 4 | |a Trangenesis marker | |
| 650 | 4 | |a Transgenesis | |
| 650 | 4 | |a Zebrafish | |
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| 700 | 1 | |a Moran, Hannah R |e verfasserin |4 aut | |
| 700 | 1 | |a Murray, Brooke F |e verfasserin |4 aut | |
| 700 | 1 | |a Scoresby, Aaron |e verfasserin |4 aut | |
| 700 | 1 | |a Klem, John R |e verfasserin |4 aut | |
| 700 | 1 | |a Eckert, Rachel L |e verfasserin |4 aut | |
| 700 | 1 | |a Lepovsky, Elizabeth |e verfasserin |4 aut | |
| 700 | 1 | |a Bertho, Sylvain |e verfasserin |4 aut | |
| 700 | 1 | |a Nieuwenhuize, Susan |e verfasserin |4 aut | |
| 700 | 1 | |a Burger, Sibylle |e verfasserin |4 aut | |
| 700 | 1 | |a D'Agati, Gianluca |e verfasserin |4 aut | |
| 700 | 1 | |a Betz, Charles |e verfasserin |4 aut | |
| 700 | 1 | |a Puller, Ann-Christin |e verfasserin |4 aut | |
| 700 | 1 | |a Felker, Anastasia |e verfasserin |4 aut | |
| 700 | 1 | |a Ditrychova, Karolina |e verfasserin |4 aut | |
| 700 | 1 | |a Bötschi, Seraina |e verfasserin |4 aut | |
| 700 | 1 | |a Affolter, Markus |e verfasserin |4 aut | |
| 700 | 1 | |a Rohner, Nicolas |e verfasserin |4 aut | |
| 700 | 1 | |a Lovely, C Ben |e verfasserin |4 aut | |
| 700 | 1 | |a Kwan, Kristen M |e verfasserin |4 aut | |
| 700 | 1 | |a Burger, Alexa |e verfasserin |4 aut | |
| 700 | 1 | |a Mosimann, Christian |e verfasserin |4 aut | |
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