Development and application of novel transgenic technology by in vivo RMCE

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This study established a targeted transgenesis system based on Cre-loxP site-specific recombination in fertilized eggs, but not in ES cells. A transgene, flanked by two mutant loxP sites, was integrated into target loci in the seed mice (containing loxP-site derivatives at Rosa26 locus) by recombinase-mediated cassette exchange (RMCE) in fertilized eggs. By applying this method, we have established a total of 25 transgenic lines showing ubiquitous, strong, and reproducible transgene expression. We also demonstrated that knockdown mice can be readily generated by PITT by taking advantage of our reproducible and highly efficient expression system.