Generation of genetically engineered mice by the piggyBac transposon system

Genetically engineered mice (GEM) are invaluable tools not only for understanding mammalian biology but also for modeling human diseases. Here we present protocols to generate GEM with the piggyBac (PB) transposon system. In the first part, we describe a transgenic procedure that co-injects the transgene carried by a PB donor plasmid and a PB transposase (PBase)-expressing helper plasmid into the pronuclei of fertilized eggs. In the second part, we provide a large-scale, cost-effective insertional mutagenesis strategy that remobilizes single-copy PB transposons in the male germ line. Given that PB can transpose in a broad spectrum of eukaryotic hosts, the protocols described here could be adapted for other species in the future.