Related Experiment Video
Updated: Jul 15, 2026
09:48
Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation
Published on: August 21, 2010
High-throughput transgenesis in Xenopus using I-SceI meganuclease
Hajime Ogino1, William B McConnell, Robert M Grainger
1Department of Biology, University of Virginia, Charlottesville, Virginia 22904, USA.
Nature Protocols
|May 10, 2007
Summary
We developed a simple and efficient Xenopus transgenesis method using I-SceI meganuclease. This technique enables high-throughput genetic studies and analysis of regulatory networks in Xenopus tropicalis.
Area of Science:
- Developmental Biology
- Genetics
- Molecular Biology
Background:
- Xenopus frogs are valuable models for studying vertebrate development and genetics.
- Efficient transgenesis methods are crucial for functional genomic studies in Xenopus.
Purpose of the Study:
- To establish a simple, efficient, and high-throughput transgenesis method for Xenopus.
- To facilitate genome-wide regulatory network studies and genetic analyses in Xenopus.
Main Methods:
- Utilizing a commercially available meganuclease (I-SceI) with a transgene construct containing its recognition sites.
- Microinjection of the I-SceI-incubated transgene construct into fertilized Xenopus eggs.
- Compatibility with simultaneous or sequential injection of mRNA or morpholino oligonucleotides.
Main Results:
- Achieved high rates of non-mosaic, promoter-dependent transgene expression (approx. 30% in X. tropicalis, 20% in X. laevis).
- Demonstrated stable transmission of transgenes from founder animals to offspring.
- Confirmed compatibility with simultaneous or sequential introduction of other reagents.
Conclusions:
- The described I-SceI-based method offers an easy, efficient, and high-throughput approach for Xenopus transgenesis.
- This technique is a powerful tool for dissecting complex regulatory networks and advancing genetic studies in Xenopus.
- The method is particularly advantageous for the rapidly breeding diploid frog, Xenopus tropicalis.
