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PiggyBac Transposon-Based Insertional Mutagenesis in Mice

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Cancer Driver Genes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1907))

Abstract

While sequencing and array-based studies are creating catalogues of genetic alterations in cancer, discriminating cancer drivers among the large sets of epigenetically, transcriptionally or posttranslationally dysregulated genes remains a challenge. Transposon-based genetic screening in mice has proven to be a powerful approach to address this challenge. Insertional mutagenesis directly flags biologically relevant genes and, combined with the transposon’s unique molecular fingerprint, facilitates the recovery of insertion sites. We have generated transgenic mouse lines harboring different versions of PiggyBac-based oncogenic transposons, which in conjunction with PiggyBac transposase mice can be used for whole-body or tissue-specific insertional mutagenesis screens. We have also developed QiSeq, a method for (semi-)quantitative transposon insertion site sequencing, which overcomes biasing limitations of previous library preparation methods. QiSeq can be used in multiplexed high-throughput formats for candidate cancer gene discovery and gives insights into the clonal distribution of insertions for the study of genetic tumor evolution.

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Author information

Authors and Affiliations

  1. The Wellcome Trust Sanger Institute, Hinxton, UK

    Mathias J. Friedrich, Iraad F. Bronner, Pentao Liu & Allan Bradley

  2. Department of Medicine II, Klinikum rechts der Isar, Technische Universität München, Munich, Germany

    Roland Rad

  3. German Cancer Research Center (DKFZ) and German Cancer Consortium (DKTK), Heidelberg, Germany

    Roland Rad

Authors
  1. Mathias J. Friedrich
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  2. Iraad F. Bronner
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  3. Pentao Liu
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  4. Allan Bradley
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  5. Roland Rad
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Corresponding author

Correspondence to Roland Rad .

Editor information

Editors and Affiliations

  1. Masonic Cancer Center, University of Minnesota Medical School, Minneapolis, MN, USA

    Timothy K. Starr

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Friedrich, M.J., Bronner, I.F., Liu, P., Bradley, A., Rad, R. (2019). PiggyBac Transposon-Based Insertional Mutagenesis in Mice. In: Starr, T. (eds) Cancer Driver Genes. Methods in Molecular Biology, vol 1907. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8967-6_14

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  • DOI: https://doi.org/10.1007/978-1-4939-8967-6_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8966-9

  • Online ISBN: 978-1-4939-8967-6

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