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Updated: Jun 23, 2026
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The Zebrafish Tol2 System: A Modular and Flexible Gateway-Based Transgenesis Approach
Published on: November 30, 2022
Simple and efficient transgenesis with meganuclease constructs in zebrafish
Daniele Soroldoni1, Benjamin M Hogan, Andrew C Oates
1Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Pfotenhauerstrasse 108, 01307, Dresden, Germany. soroldon@mpi-cbg.de
Methods in Molecular Biology (Clifton, N.J.)
|April 21, 2009
Summary
Generating transgenic zebrafish is now more efficient using I-SceI meganuclease. This method improves transgene integration and increases transgenesis success rates, simplifying genetic experiments.
Area of Science:
- Genetics
- Molecular Biology
- Developmental Biology
Background:
- Traditional zebrafish transgenesis via microinjection has limitations.
- These include mosaic transgene distribution and low efficiency.
Purpose of the Study:
- To optimize a protocol for I-SceI meganuclease-mediated transgenesis in zebrafish.
- To enhance the efficiency and reliability of generating transgenic zebrafish lines.
Main Methods:
- Utilized I-SceI meganuclease with reporter constructs flanked by I-SceI sites.
- Optimized a protocol for coinjection into early zebrafish embryos.
- Assessed reporter gene expression in F0 generation embryos.
Main Results:
- Achieved earlier transgene integration into the host genome.
- Increased transgenesis frequencies up to 45%, compared to 5% without I-SceI.
- Demonstrated reliable transient reporter expression in F0 embryos.
Conclusions:
- The optimized I-SceI protocol significantly improves transgenic line generation in zebrafish.
- This method is a powerful tool for various genetic applications, including cell tracing and labeling.
