Zuber Michael E, Nihart Heather S, Zhuo Xinming, Babu Sudha, Knox Barry E
Department of Ophthalmology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, NY 13210, USA.
Genesis. 2012 Mar;50(3):325-32. doi: 10.1002/dvg.22006. Epub 2012 Feb 15.
Transgenesis is an essential, powerful tool for investigating gene function and the activities of enhancers, promoters, and transcription factors in the chromatin environment. In Xenopus, current methods generate germ-line transgenics by random insertion, often resulting in mosaicism, position-dependent variations in expression, and lab-to-lab differences in efficiency. We have developed and tested a Xenopus FLP-FRT recombinase-mediated transgenesis (X-FRMT) method. We demonstrate transgenesis of Xenopus laevis by FLP-catalyzed recombination of donor plasmid cassettes into F(1) tadpoles with host cassette transgenes. X-FRMT provides a new method for generating transgenic Xenopus. Once Xenopus lines harboring single host cassettes are generated, X-FRMT should allow for the targeting of transgenes to well-characterized integration site(s), requiring no more special reagents or training than that already common to most Xenopus labs.
转基因技术是研究基因功能以及染色质环境中增强子、启动子和转录因子活性的一种重要且强大的工具。在非洲爪蟾中，目前的方法通过随机插入产生种系转基因，这常常导致镶嵌性、表达的位置依赖性变异以及不同实验室间效率的差异。我们开发并测试了一种非洲爪蟾FLP-FRT重组酶介导的转基因方法（X-FRMT）。我们通过FLP催化供体质粒盒与带有宿主盒转基因的F(1)蝌蚪进行重组，展示了非洲爪蟾的转基因过程。X-FRMT为产生转基因非洲爪蟾提供了一种新方法。一旦产生了携带单个宿主盒的非洲爪蟾品系，X-FRMT应该能够将转基因靶向到特征明确的整合位点，所需的特殊试剂或培训不会比大多数非洲爪蟾实验室常用的更多。
