Abstract
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.
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Acknowledgement
This work was supported by Chinese Key Projects for Basic Research (973) grant 2006CB806701, Hi-tech Research and Development Project (863) grant 2007AA022101, National Natural Science Foundation of China (NSFC) grants 30550006 and 81228019, Shanghai Rising-Star Program grant 10QH1400100, and the “Shu Guang” project grant 09SG04 of Shanghai Municipal Education Commission and Shanghai Education Development Foundation.
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Ding, S., Xu, T., Wu, X. (2014). Generation of Genetically Engineered Mice by the piggyBac Transposon System. In: Singh, S., Coppola, V. (eds) Mouse Genetics. Methods in Molecular Biology, vol 1194. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1215-5_9
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DOI: https://doi.org/10.1007/978-1-4939-1215-5_9
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