Generation of Mouse Models of Cancer Using Transposon‐Mediated Approaches


Cancer genomes harbour a formidable genetic heterogeneity that makes determining the genes involved in tumour development or therapeutic response challenging. Deoxyribonucleic acid (DNA) transposon systems have enabled the development of a new generation of mouse models to help tackle this complex situation. These transposon‐based genetically engineered mouse models of cancer have provided a powerful tool for cancer gene discovery, complementing genomic studies in human tumour specimens that have been accomplished during the past 15 years. In addition, the high gene delivery efficiency of DNA transposons has been used to generate reverse genetic mouse models of cancer in order to study the function of specific cancer genes. Overall, DNA transposons have reinforced and advanced the study of cancer pathogenesis, unveiling cancer promoting mechanisms and potential therapeutic targets.

Key Concepts

  • Cancer genomes harbour a complex genetic landscape.
  • Mouse models of cancer have extensively contributed to the understanding of the tumour pathogenesis.
  • Transposon‐based mouse models of cancer provide a powerful tool for cancer gene discovery.
  • Sleeping Beauty and piggyBac transposon systems are complementary.
  • DNA transposons allow the functional validation of cancer genes in vivo.

Keywords: DNA transposon; Sleeping Beauty; piggyBac; genetically engineered mouse model; cancer genetics; insertional mutagenesis; reverse genetics; gene discovery

Figure 1. DNA transposon system. (a) Transposon structure. IR/DR, inverted repeat/direct repeat sequence; SA, splice acceptor; pA, polyadenylation sequence and SD, splice donor. (b) Effects of transposons on target genes. A hypothetical gene is shown in grey with a promoter (arrow) and three exons (boxes). When transposons are mobilised and integrated in an intron, they can either trap upstream exons inactivating potential tumour suppressor genes (loss‐of‐function mutations) or activate the expression of potential downstream proto‐oncogenes or dominant‐negative forms of tumour suppressor genes (gain‐of‐function mutations). (c) Insertional mutagenesis screen. A transposon concatemer (red rectangle) is located in a mouse chromosome. In the presence of the transposase (green ellipse), transposons are mobilised from the donor concatemer and reintegrated throughout the genome using a ‘cut‐and‐paste′ mechanism.
Figure 2. Constitutive mutagenesis. (a) Constitutive transposase allele. Whole‐body transposase expression is driven by a constitutive promoter (CP). (b) Crossing strategy for the generation of constitutive transposon‐mediated GEMMs of cancer.
Figure 3. Tissue‐specific mutagenesis. (a) Conditional Rosa26‐LSL‐SB11 allele. Cre recombinase expression from a tissue‐specific promoter (TSP) leads to the excision of the LoxP‐STOP‐LoxP cassette, allowing SB11 expression in the tissue‐of‐interest. Black triangle: LoxP site. (b) Crossing strategy for the generation of transposon‐mediated GEMMs of cancer using conditional transposase strains. (c) Tissue‐specific transposase allele. Transposase expression is driven by a TSP. (d) Crossing strategy for the generation of transposon‐mediated GEMMs of cancer using tissue‐specific transposase strains.
Figure 4. Reverse genetic for cancer gene validation. (a) DNA transposons harbouring selected cDNAs and/or shRNAs can be delivered in the tissue of interest of transposase‐expressing mice (green circle). (b) DNA transposons harbouring selected cDNAs and/or shRNAs can be codelivered together with transposase‐expressing vectors in mice that do not harbour a transposase expressing allele. IR/DR, inverted repeat/direct repeat sequence and pA, polyadenylation signal.


Bard‐Chapeau EA, Nguyen AT, Rust AG, et al. (2014) Transposon mutagenesis identifies genes driving hepatocellular carcinoma in a chronic hepatitis B mouse model. Nature Genetics 46: 24–32.

Bergemann TL, Starr TK, Yu H, et al. (2012) New methods for finding common insertion sites and co‐occurring common insertion sites in transposon‐ and virus‐based genetic screens. Nucleic Acids Research 40: 3822–3833.

Bermejo‐Rodriguez C and Perez‐Mancera PA (2015) Use of DNA transposons for functional genetic screens in mouse models of cancer. Current Opinion in Biotechnology 35: 103–110.

Bos JL (1989) ras oncogenes in human cancer: a review. Cancer Research 49: 4682–4689.

Carlson CM, Dupuy AJ, Fritz S, et al. (2003) Transposon mutagenesis of the mouse germline. Genetics 165: 243–256.

Carlson CM, Frandsen JL, Kirchhof N, McIvor RS and Largaespada DA (2005) Somatic integration of an oncogene‐harboring Sleeping Beauty transposon models liver tumor development in the mouse. Proceedings of the National Academy of Sciences of the United States of America 102: 17059–17064.

Collier LS, Carlson CM, Ravimohan S, Dupuy AJ and Largaespada DA (2005) Cancer gene discovery in solid tumours using transposon‐based somatic mutagenesis in the mouse. Nature 436: 272–276.

Collier LS, Adams DJ, Hackett CS, et al. (2009) Whole‐body sleeping Beauty mutagenesis can cause penetrant leukemia/lymphoma and rare high‐grade glioma without associated embryonic lethality. Cancer Research 69: 8429–8437.

Copeland NG and Jenkins NA (2010) Harnessing transposons for cancer gene discovery. Nature Reviews Cancer 10: 696–706.

de Jong J, Akhtar W, Badhai J, et al. (2014) Chromatin landscapes of retroviral and transposon integration profiles. PLoS Genetics 10: e1004250.

de Ridder J, Uren A, Kool J, Reinders M and Wessels L (2006) Detecting statistically significant common insertion sites in retroviral insertional mutagenesis screens. PLoS Computational Biology 2: e166.

Ding S, Wu X, Li G, et al. (2005) Efficient transposition of the piggyBac (PB) transposon in mammalian cells and mice. Cell 122: 473–483.

Dupuy AJ, Akagi K, Largaespada DA, Copeland NG and Jenkins NA (2005) Mammalian mutagenesis using a highly mobile somatic Sleeping Beauty transposon system. Nature 436: 221–226.

Dupuy AJ, Rogers LM, Kim J, et al. (2009) A modified Sleeping Beauty transposon system that can be used to model a wide variety of human cancers in mice. Cancer Research 69: 8150–8156.

Fearon ER and Vogelstein B (1990) A genetic model for colorectal tumorigenesis. Cell 61: 759–767.

Fraser MJ, Ciszczon T, Elick T and Bauser C (1996) Precise excision of TTAA‐specific lepidopteran transposons piggyBac (IFP2) and tagalong (TFP3) from the baculovirus genome in cell lines from two species of Lepidoptera. Insect Molecular Biology 5: 141–151.

Frese KK and Tuveson DA (2007) Maximizing mouse cancer models. Nature Reviews Cancer 7: 645–658.

Geurts AM, Yang Y, Clark KJ, et al. (2003) Gene transfer into genomes of human cells by the Sleeping Beauty transposon system. Molecular Therapy 8: 108–117.

Ivics Z, Hackett PB, Plasterk RH and Izsvak Z (1997) Molecular reconstruction of Sleeping Beauty, a Tc1‐like transposon from fish, and its transposition in human cells. Cell 91: 501–510.

Kang TW, Yevsa T, Woller N, et al. (2011) Senescence surveillance of pre‐malignant hepatocytes limits liver cancer development. Nature 479: 547–551.

Keng VW, Villanueva A, Chiang DY, et al. (2009) A conditional transposon‐based insertional mutagenesis screen for genes associated with mouse hepatocellular carcinoma. Nature Biotechnology 27: 264–274.

Keng VW, Tschida BR, Bell JB and Largaespada DA (2011) Modeling hepatitis B virus X‐induced hepatocellular carcinoma in mice with the Sleeping Beauty transposon system. Hepatology 53: 781–790.

Keng VW, Sia D, Sarver AL, et al. (2013) Sex bias occurrence of hepatocellular carcinoma in Poly7 molecular subclass is associated with EGFR. Hepatology 57: 120–130.

Li MA, Turner DJ, Ning Z, et al. (2011) Mobilization of giant piggyBac transposons in the mouse genome. Nucleic Acids Research 39: e148.

Li MA, Pettitt SJ, Eckert S, et al. (2013) The piggyBac transposon displays local and distant reintegration preferences and can cause mutations at noncanonical integration sites. Molecular and Cellular Biology 33: 1317–1330.

Mann KM, Ward JM, Yew CC, et al. (2012) Sleeping Beauty mutagenesis reveals cooperating mutations and pathways in pancreatic adenocarcinoma. Proceedings of the National Academy of Sciences of the United States of America 109: 5934–5941.

March HN, Rust AG, Wright NA, et al. (2011) Insertional mutagenesis identifies multiple networks of cooperating genes driving intestinal tumorigenesis. Nature Genetics 43: 1202–1209.

Mates L, Chuah MK, Belay E, et al. (2009) Molecular evolution of a novel hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates. Nature Genetics 41: 753–761.

Narod SA (2002) Modifiers of risk of hereditary breast and ovarian cancer. Nature Reviews Cancer 2: 113–123.

Perez‐Mancera PA, Rust AG, van der Weyden L, et al. (2012) The deubiquitinase USP9X suppresses pancreatic ductal adenocarcinoma. Nature 486: 266–270.

Perna D, Karreth FA, Rust AG, et al. (2015) BRAF inhibitor resistance mediated by the AKT pathway in an oncogenic BRAF mouse melanoma model. Proceedings of the National Academy of Sciences of the United States of America 112: E536–E545.

Platz A, Egyhazi S, Ringborg U and Hansson J (2008) Human cutaneous melanoma; a review of NRAS and BRAF mutation frequencies in relation to histogenetic subclass and body site. Molecular Oncology 1: 395–405.

Rad R, Rad L, Wang W, et al. (2010) PiggyBac transposon mutagenesis: a tool for cancer gene discovery in mice. Science 330: 1104–1107.

Rad R, Rad L, Wang W, et al. (2015) A conditional piggyBac transposition system for genetic screening in mice identifies oncogenic networks in pancreatic cancer. Nature Genetics 47: 47–56.

Riordan JD, Keng VW, Tschida BR, et al. (2013) Identification of rtl1, a retrotransposon‐derived imprinted gene, as a novel driver of hepatocarcinogenesis. PLoS Genetics 9: e1003441.

Rudalska R, Dauch D, Longerich T, et al. (2014) In vivo RNAi screening identifies a mechanism of sorafenib resistance in liver cancer. Nature Medicine 20: 1138–1146.

Sinzelle L, Izsvak Z and Ivics Z (2009) Molecular domestication of transposable elements: from detrimental parasites to useful host genes. Cellular and Molecular Life Sciences 66: 1073–1093.

Starr TK, Allaei R, Silverstein KA, et al. (2009) A transposon‐based genetic screen in mice identifies genes altered in colorectal cancer. Science 323: 1747–1750.

Starr TK, Scott PM, Marsh BM, et al. (2011) A Sleeping Beauty transposon‐mediated screen identifies murine susceptibility genes for adenomatous polyposis coli (Apc)‐dependent intestinal tumorigenesis. Proceedings of the National Academy of Sciences of the United States of America 108: 5765–5770.

Stratton MR, Campbell PJ and Futreal PA (2009) The cancer genome. Nature 458: 719–724.

Takeda H, Wei Z, Koso H, et al. (2015) Transposon mutagenesis identifies genes and evolutionary forces driving gastrointestinal tract tumor progression. Nature Genetics 47: 142–150.

Uren AG, Kool J, Berns A and van Lohuizen M (2005) Retroviral insertional mutagenesis: past, present and future. Oncogene 24: 7656–7672.

Uren AG, Mikkers H, Kool J, et al. (2009) A high‐throughput splinkerette‐PCR method for the isolation and sequencing of retroviral insertion sites. Nature Protocols 4: 789–798.

van der Weyden L, Giotopoulos G, Rust AG, et al. (2011) Modeling the evolution of ETV6‐RUNX1‐induced B‐cell precursor acute lymphoblastic leukemia in mice. Blood 118: 1041–1051.

Wang W, Lin C, Lu D, et al. (2008) Chromosomal transposition of PiggyBac in mouse embryonic stem cells. Proceedings of the National Academy of Sciences of the United States of America 105: 9290–9295.

Wangensteen KJ, Wilber A, Keng VW, et al. (2008) A facile method for somatic, lifelong manipulation of multiple genes in the mouse liver. Hepatology 47: 1714–1724.

Wiesner SM, Decker SA, Larson JD, et al. (2009) De novo induction of genetically engineered brain tumors in mice using plasmid DNA. Cancer Research 69: 431–439.

Wilson MH, Coates CJ and George AL Jr (2007) PiggyBac transposon‐mediated gene transfer in human cells. Molecular Therapy 15: 139–145.

Wu X, Northcott PA, Dubuc A, et al. (2012) Clonal selection drives genetic divergence of metastatic medulloblastoma. Nature 482: 529–533.

Yant SR, Huang Y, Akache B and Kay MA (2007) Site‐directed transposon integration in human cells. Nucleic Acids Research 35: e50.

Zayed H, Izsvak Z, Walisko O and Ivics Z (2004) Development of hyperactive Sleeping Beauty transposon vectors by mutational analysis. Molecular Therapy 9: 292–304.

Further Reading

Been RA, Linden MA, Hager CJ, et al. (2014) Genetic signature of histiocytic sarcoma revealed by a Sleeping Beauty transposon genetic screen in mice. PLoS One 9: e97280.

Berquam‐Vrieze KE, Nannapaneni K, Brett BT, et al. (2011) Cell of origin strongly influences genetic selection in a mouse model of T‐ALL. Blood 118: 4646–4656.

Howell VM and Colvin EK (2014) Genetically engineered insertional mutagenesis in mice to model cancer: Sleeping Beauty. Methods in Molecular Biology 1194: 367–383.

Mann KM, Jenkins NA, Copeland NG and Mann MB (2014) Transposon insertional mutagenesis models of cancer. Cold Spring Harbor Protocols 2014: 235–247.

Mann MB, Black MA, Jones DJ, et al. (2015) Transposon mutagenesis identifies genetic drivers of Braf(V600E) melanoma. Nature Genetics 47: 486–495.

Moriarity BS, Otto GM, Rahrmann EP, et al. (2015) A Sleeping Beauty forward genetic screen identifies new genes and pathways driving osteosarcoma development and metastasis. Nature Genetics 47: 615–624.

Quintana RM, Dupuy AJ, Bravo A, et al. (2013) A transposon‐based analysis of gene mutations related to skin cancer development. Journal of Investigative Dermatology 133: 239–248.

Rahrmann EP, Watson AL, Keng VW, et al. (2013) Forward genetic screen for malignant peripheral nerve sheath tumor formation identifies new genes and pathways driving tumorigenesis. Nature Genetics 45: 756–766.

Tschida BR, Largaespada DA and Keng VW (2014) Mouse models of cancer: Sleeping Beauty transposons for insertional mutagenesis screens and reverse genetic studies. Seminars in Cell and Developmental Biology 27: 86–95.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
Bermejo‐Rodríguez, Camino, and Pérez‐Mancera, Pedro A(Mar 2017) Generation of Mouse Models of Cancer Using Transposon‐Mediated Approaches. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0026891]