Site-specific integrase-mediated transgenesis in mice via pronuclear injection

• DOI: 10.1073/pnas.1019507108
• Publication Date: 2011-04-05T03:19:19Z
• Authors: Bosiljka Tasic; Simon Hippenmeyer; Charlene Wang; Matthew Gamboa; Hui Zong; Yanru Chen-Tsai; Liqun Luo
• Keywords: Transgenesis; Pronucleus; Bacterial artificial chromosome; Recombineering; Site-specific recombination

Microinjection of recombinant DNA into zygotic pronuclei has been widely used for producing transgenic mice. However, with this method, the insertion site, integrity, and copy number transgene cannot be controlled. Here, we present an integrase-based approach to produce mice via pronuclear injection, whereby intact single-copy can inserted predetermined chromosomal loci high efficiency (up 40%), faithfully transmitted through generations. We show that neighboring elements bacterial within cause profound silencing expression variability marker. Removal these undesirable leads global high-level marker from transgenes driven by a ubiquitous promoter. also obtained faithful tissue-specific The technique presented here will greatly facilitate murine transgenesis precise structure/function dissection mammalian gene function regulation in vivo.