Generation of Mouse Models of Cancer Using Transposon‐Mediated Approaches

Abstract

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.
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Further Reading

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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. http://www.els.net [doi: 10.1002/9780470015902.a0026891]