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Generation of Genetically Engineered Mice by the piggyBac Transposon System

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Mouse Genetics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1194))

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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Authors and Affiliations

  1. State Key Laboratory of Genetic Engineering and Institute of Developmental Biology and Molecular Medicine, Fudan-Yale Biomedical Research Center, School of Life Sciences, Fudan University, Shanghai, 200433, China

    Sheng Ding, Tian Xu & Xiaohui Wu

  2. Howard Hughes Medical Institute, Department of Genetics, Yale University School of Medicine, New Haven, CT, 06536, USA

    Sheng Ding & Tian Xu

Authors
  1. Sheng Ding
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  2. Tian Xu
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  3. Xiaohui Wu
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Corresponding author

Correspondence to Xiaohui Wu .

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Editors and Affiliations

  1. Stem Cell Regulation and Animal Aging Section, Basic Research Laboratory,, National Cancer Institute, National Institutes of Health, Frederick, MD, USA

    Shree Ram Singh

  2. Genetically Engineered Mouse Modeling Core/Department of Molecular Virology, Immunology and Medical Genetics, Wexner Medical Center, Comprehensive Cancer Center, The Ohio State University, Columbus, OH, USA

    Vincenzo Coppola

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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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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1214-8

  • Online ISBN: 978-1-4939-1215-5

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