Ogino Hajime, McConnell William B, Grainger Robert M
Department of Biology, University of Virginia, Charlottesville, Virginia 22904, USA.
Nat Protoc. 2006;1(4):1703-10. doi: 10.1038/nprot.2006.208.
In this report we describe an easy, highly efficient transgenesis method for Xenopus. The method is very simple; a commercially available meganuclease, I-SceI, is incubated with a transgene construct carrying its recognition sites, and is subsequently microinjected into fertilized eggs. Approximately 30% (in Xenopus tropicalis) or 20% (in Xenopus laevis) of injected embryos exhibit non-mosaic, promoter-dependent transgene expression, and transgenes from the founder animals are transmitted to offspring. The method is compatible with mRNA or antisense morpholino oligonucleotide injection, and these secondary reagents can be introduced simultaneously or sequentially with a transgene to test their interaction. This high-throughput transgenic technique will be a powerful tool for studying the complex wiring of regulatory networks at the genome-wide level, as well as for facilitating genetic studies in the rapidly breeding diploid frog, X. tropicalis.
在本报告中，我们描述了一种用于非洲爪蟾的简便、高效的转基因方法。该方法非常简单；将一种市售的巨核酸酶I-SceI与携带其识别位点的转基因构建体一起孵育，随后显微注射到受精卵中。大约30%（热带爪蟾）或20%（非洲爪蟾）的注射胚胎表现出非镶嵌、启动子依赖性转基因表达，并且来自奠基动物的转基因可传递给后代。该方法与mRNA或反义吗啉代寡核苷酸注射兼容，并且这些辅助试剂可以与转基因同时或相继导入，以测试它们之间的相互作用。这种高通量转基因技术将成为在全基因组水平研究调控网络复杂线路的有力工具，同时也有助于在快速繁殖的二倍体青蛙热带爪蟾中开展遗传学研究。
