Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Review Article
  • Published:

Insertional mutagenesis in mice: new perspectives and tools

Nature Reviews Genetics volume 6, pages 568–580 (2005)Cite this article

Key Points

  • Recent initiatives have outlined the strategies and goals of the mouse community to achieve complete functional annotation of the mouse genome.

  • Insertional mutagenesis strategies have proved invaluable for mouse genetics research, and new technologies aim to improve the usefulness of this approach.

  • The Velocigene system, developed by Regeneron Pharmaceuticals, uses a BAC recombination system to generate targeted knockouts faster and more efficiently than traditional gene targeting systems.

  • The mutagenic and chromosome engineering resource (MICER) is a resource of insertion vectors that can target specified genes for mutation with great efficiency and can allow the production of templates for chromosome engineering.

  • The progress of genome-wide gene-trap screens has been substantial, and the advent of expression screening allows for prescreening in culture for gene classes of interest.

  • Using the Cre/loxP system or exploiting the increased incidence of non-sister chromatid exchange in Bloom syndrome cells allows the induction of mitotic recombination in embryonic stem cells. These approaches can be used to generate biallelic homozygous mutations in culture for recessive genetic screens.

  • The optimization of several transposable elements has led to the development of efficient systems for in vivo transposon mutagenesis for forward genetic screens. These elements can also potentially be used for generating substrates for chromosome engineering, gain-of-function screens and somatic cell insertional mutagenesis in the mouse.

  • A synthesis of insertional mutagenesis technologies will be necessary to achieve complete genome coverage for functional annotation, and will compliment alternative approaches including chemical mutagenesis.

Abstract

Insertional mutagenesis has been at the core of functional genomics in many species. In the mouse, improved vectors and methodologies allow easier genome-wide and phenotype-driven insertional mutagenesis screens. The ability to generate homozygous diploid mutations in mouse embryonic stem cells allows prescreening for specific null phenotypes prior to in vivo analysis. In addition, the discovery of active transposable elements in vertebrates, and their development as genetic tools, has led to in vivo forward insertional mutagenesis screens in the mouse. These new technologies will greatly contribute to the speed and ease with which we achieve complete functional annotation of the mouse genome.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Insertional mutagenesis approaches and mechanisms.
Figure 2: Expression trapping in embryonic stem cells.
Figure 3: Induced mitotic recombination using Cre/loxP or a Bloom syndrome mutant background.
Figure 4: Exploiting the 'local hopping' of the Sleeping Beauty transposon.

Similar content being viewed by others

References

  1. Nadeau, J. H. et al. Sequence interpretation. Functional annotation of mouse genome sequences. Science 291, 1251–1255 (2001).

    Article  CAS  PubMed  Google Scholar 

  2. Austin, C. P. et al. The knockout mouse project. Nature Genet. 36, 921–924 (2004).

    Article  CAS  PubMed  Google Scholar 

  3. Auwerx, J. et al. The European dimension for the mouse genome mutagenesis program. Nature Genet. 36, 925–927 (2004). References 1–3 outline the goals and approaches to be used to achieve complete functional annotation of the mouse genome through mutagenesis and phenotype analysis. Reference 1 describes the establishment of the IMMC, and references 2 and 3 are a further refinement of approaches and goals that reflect the thoughts of the mouse community today.

    Article  CAS  PubMed  Google Scholar 

  4. Justice, M. J., Noveroske, J. K., Weber, J. S., Zheng, B. & Bradley, A. Mouse ENU mutagenesis. Hum. Mol. Genet. 8, 1955–1963 (1999).

    Article  CAS  PubMed  Google Scholar 

  5. Hrabe de Angelis, M. H. et al. Genome-wide, large-scale production of mutant mice by ENU mutagenesis. Nature Genet. 25, 444–447 (2000).

    Article  CAS  PubMed  Google Scholar 

  6. Nolan, P. M. et al. A systematic, genome-wide, phenotype-driven mutagenesis programme for gene function studies in the mouse. Nature Genet. 25, 440–443 (2000).

    Article  CAS  PubMed  Google Scholar 

  7. Yang, X. W., Model, P. & Heintz, N. Homologous recombination based modification in Escherichia coli and germline transmission in transgenic mice of a bacterial artificial chromosome. Nature Biotechnol. 15, 859–865 (1997).

    Article  CAS  Google Scholar 

  8. Yu, D. et al. An efficient recombination system for chromosome engineering in Escherichia coli. Proc. Natl Acad. Sci. USA 97, 5978–5983 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Testa, G. et al. Engineering the mouse genome with bacterial artificial chromosomes to create multipurpose alleles. Nature Biotechnol. 21, 443–447 (2003).

    Article  CAS  Google Scholar 

  10. Yang, Y. & Seed, B. Site-specific gene targeting in mouse embryonic stem cells with intact bacterial artificial chromosomes. Nature Biotechnol. 21, 447–451 (2003).

    Article  CAS  Google Scholar 

  11. Valenzuela, D. M. et al. High-throughput engineering of the mouse genome coupled with high-resolution expression analysis. Nature Biotechnol. 21, 652–659 (2003).

    Article  CAS  Google Scholar 

  12. Adams, D. J. et al. Mutagenic insertion and chromosome engineering resource (MICER). Nature Genet. 36, 867–871 (2004). This paper describes the MICER system with regard to application, genome coverage and availability of insertion vectors for targeted mutagenesis in mouse ES cells.

    Article  CAS  PubMed  Google Scholar 

  13. Zheng, B., Mills, A. A. & Bradley, A. A system for rapid generation of coat color-tagged knockouts and defined chromosomal rearrangements in mice. Nucleic Acids Res. 27, 2354–2360 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Yu, Y. & Bradley, A. Engineering chromosomal rearrangements in mice. Nature Rev. Genet. 2, 780–790 (2001).

    Article  CAS  PubMed  Google Scholar 

  15. Durick, K., Mendlein, J. & Xanthopoulos, K. G. Hunting with traps: genome-wide strategies for gene discovery and functional analysis. Genome Res. 9, 1019–1025 (1999).

    Article  CAS  PubMed  Google Scholar 

  16. Stanford, W. L., Cohn, J. B. & Cordes, S. P. Gene-trap mutagenesis: past, present and beyond. Nature Rev. Genet. 2, 756–768 (2001).

    Article  CAS  PubMed  Google Scholar 

  17. Skarnes, W. C. et al. A public gene trap resource for mouse functional genomics. Nature Genet. 36, 543–544 (2004).

    Article  CAS  PubMed  Google Scholar 

  18. Zambrowicz, B. P. et al. Disruption and sequence identification of 2,000 genes in mouse embryonic stem cells. Nature 392, 608–611 (1998). This paper describes the efforts of Lexicon Genetics to create an ES cell library that contains gene-trap insertions within every gene of the mouse. This work used a combined promoter-trap/poly(A)-trap vector to identify genes.

    Article  CAS  PubMed  Google Scholar 

  19. Stryke, D. et al. BayGenomics: a resource of insertional mutations in mouse embryonic stem cells. Nucleic Acids Res. 31, 278–281 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Zambrowicz, B. P. et al. Wnk1 kinase deficiency lowers blood pressure in mice: a gene-trap screen to identify potential targets for therapeutic intervention. Proc. Natl Acad. Sci. USA 100, 14109–14114 (2003).

    Article  PubMed  PubMed Central  Google Scholar 

  21. Shigeoka, T., Kawaichi, M. & Ishida, Y. Suppression of nonsense-mediated mRNA decay permits unbiased gene trapping in mouse embryonic stem cells. Nucleic Acids Res. 33, e20 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Hansen, J. et al. A large-scale, gene-driven mutagenesis approach for the functional analysis of the mouse genome. Proc. Natl Acad. Sci. USA 100, 9918–9922 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Bushman, F. D. Targeting survival: integration site selection by retroviruses and LTR-retrotransposons. Cell 115, 135–138 (2003).

    Article  CAS  PubMed  Google Scholar 

  24. Skarnes, W. C., Moss, J. E., Hurtley, S. M. & Beddington, R. S. Capturing genes encoding membrane and secreted proteins important for mouse development. Proc. Natl Acad. Sci. USA 92, 6592–6596 (1995).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Mitchell, K. J. et al. Functional analysis of secreted and transmembrane proteins critical to mouse development. Nature Genet. 28, 241–249 (2001). This paper describes the use of a specialized gene-trap vector designed to trap exclusively genes that encode secreted and transmembrane proteins. It reports the characterization of one of the largest collections of mouse mutants elicited in ES cells using gene traps.

    Article  CAS  PubMed  Google Scholar 

  26. Leighton, P. A. et al. Defining brain wiring patterns and mechanisms through gene trapping in mice. Nature 410, 174–179 (2001).

    Article  CAS  PubMed  Google Scholar 

  27. Zhang, W. et al. The functional landscape of mouse gene expression. J. Biol. 3, 21 (2004).

  28. Hirashima, M., Bernstein, A., Stanford, W. L. & Rossant, J. Gene-trap expression screening to identify endothelial-specific genes. Blood 104, 711–718 (2004).

    Article  CAS  PubMed  Google Scholar 

  29. Medico, E., Gambarotta, G., Gentile, A., Comoglio, P. M. & Soriano, P. A gene trap vector system for identifying transcriptionally responsive genes. Nature Biotechnol. 19, 579–582 (2001).

    Article  CAS  Google Scholar 

  30. Chen, Y. T., Liu, P. & Bradley, A. Inducible gene trapping with drug-selectable markers and Cre/loxP to identify developmentally regulated genes. Mol. Cell Biol. 24, 9930–9941 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Chen, W. V., Delrow, J., Corrin, P. D., Frazier, J. P. & Soriano, P. Identification and validation of PDGF transcriptional targets by microarray-coupled gene-trap mutagenesis. Nature Genet 36, 304–312 (2004). This paper describes the coupling of gene-trap technology to microarray analysis. RACE products generated from gene-trap insertions of an ES cell library are spotted on a microarray to identify genes transcriptionally responsive to platelet-derived growth factor in wild-type or mutant cells. This allows identification of ES cell clones harbouring mutations in such genes.

    Article  CAS  PubMed  Google Scholar 

  32. Harrington, J. J. et al. Creation of genome-wide protein expression libraries using random activation of gene expression. Nature Biotechnol. 19, 440–445 (2001).

    Article  Google Scholar 

  33. Pellegrini, S., John, J., Shearer, M., Kerr, I. M. & Stark, G. R. Use of a selectable marker regulated by αinterferon to obtain mutations in the signaling pathway. Mol. Cell. Biol. 9, 4605–4612 (1989).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Zheng, B., Sage, M., Sheppeard, E. A., Jurecic, V. & Bradley, A. Engineering mouse chromosomes with Cre-loxP : range, efficiency, and somatic applications. Mol. Cell. Biol. 20, 648–655 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Koike, H. et al. Efficient biallelic mutagenesis with Cre/loxP-mediated inter-chromosomal recombination. EMBO Rep. 3, 433–437 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Liu, P., Jenkins, N. A. & Copeland, N. G. Efficient Cre-loxP-induced mitotic recombination in mouse embryonic stem cells. Nature Genet. 30, 66–72 (2002). References 35 and 36 are the first to report the induction of mitotic recombination in embryonic stem cells using the Cre/ loxP system to elicit homozygous mutations in culture.

    Article  CAS  PubMed  Google Scholar 

  37. Smith, A. J. et al. A site-directed chromosomal translocation induced in embryonic stem cells by Cre-loxP recombination. Nature Genet. 9, 376–385 (1995).

    Article  CAS  PubMed  Google Scholar 

  38. Ellis, N. A. et al. The Bloom's syndrome gene product is homologous to RecQ helicases. Cell 83, 655–666 (1995).

    Article  CAS  PubMed  Google Scholar 

  39. German, J. Bloom syndrome: a mendelian prototype of somatic mutational disease. Medicine (Baltimore) 72, 393–406 (1993).

    Article  CAS  Google Scholar 

  40. Groden, J., Nakamura, Y. & German, J. Molecular evidence that homologous recombination occurs in proliferating human somatic cells. Proc. Natl Acad. Sci. USA 87, 4315–4319 (1990).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Wu, L. & Hickson, I. D. The Bloom's syndrome helicase suppresses crossing over during homologous recombination. Nature 426, 870–874 (2003).

    Article  CAS  PubMed  Google Scholar 

  42. Chester, N., Kuo, F., Kozak, C., O'Hara, C. D. & Leder, P. Stage-specific apoptosis, developmental delay, and embryonic lethality in mice homozygous for a targeted disruption in the murine Bloom's syndrome gene. Genes Dev. 12, 3382–33893 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Goss, K. H. et al. Enhanced tumor formation in mice heterozygous for Blm mutation. Science 297, 2051–2053 (2002).

    Article  CAS  PubMed  Google Scholar 

  44. Luo, G. et al. Cancer predisposition caused by elevated mitotic recombination in Bloom mice. Nature Genet. 26, 424–429 (2000).

    Article  CAS  PubMed  Google Scholar 

  45. Guo, G., Wang, W. & Bradley, A. Mismatch repair genes identified using genetic screens in Blm-deficient embryonic stem cells. Nature 429, 891–895 (2004).

    Article  CAS  PubMed  Google Scholar 

  46. Yusa, K. et al. Genome-wide phenotype analysis in ES cells by regulated disruption of Bloom's syndrome gene. Nature 429, 896–899 (2004). References 45 and 46 were the first to exploit the increased incidence of somatic homologous recombination of cells deficient of the Bloom's RecQ helicase to produce biallelic mutations in diploid mouse ES cells.

    Article  CAS  PubMed  Google Scholar 

  47. Zwaal, R. R., Broeks, A., van Meurs, J., Groenen, J. T. & Plasterk, R. H. Target-selected gene inactivation in Caenorhabditis elegans by using a frozen transposon insertion mutant bank. Proc. Natl Acad. Sci. USA 90, 7431–7435 (1993).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Ross-Macdonald, P. et al. Large-scale analysis of the yeast genome by transposon tagging and gene disruption. Nature 402, 413–418 (1999).

    Article  CAS  PubMed  Google Scholar 

  49. Schmidt, R. J., Burr, F. A. & Burr, B. Transposon tagging and molecular analysis of the maize regulatory locus opaque-2. Science 238, 960–963 (1987).

    Article  CAS  PubMed  Google Scholar 

  50. Spradling, A. C. et al. Gene disruptions using P transposable elements: an integral component of the Drosophila genome project. Proc. Natl Acad. Sci. USA 92, 10824–10830 (1995).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Bellen, H. J. et al. P-element-mediated enhancer detection: a versatile method to study development in Drosophila. Genes Dev. 3, 1288–1300 (1989).

    Article  CAS  PubMed  Google Scholar 

  52. Bier, E. et al. Searching for pattern and mutation in the Drosophila genome with a P-lacZ vector. Genes Dev. 3, 1273–1287 (1989).

    Article  CAS  PubMed  Google Scholar 

  53. Wilson, C. et al. P-element-mediated enhancer detection: an efficient method for isolating and characterizing developmentally regulated genes in Drosophila. Genes Dev. 3, 1301–1313 (1989).

    Article  CAS  PubMed  Google Scholar 

  54. Cooley, L., Kelley, R. & Spradling, A. Insertional mutagenesis of the Drosophila genome with single P elements. Science 239, 1121–1128 (1988).

    Article  CAS  PubMed  Google Scholar 

  55. Rorth, P. et al. Systematic gain-of-function genetics in Drosophila. Development 125, 1049–10457 (1998).

    CAS  PubMed  Google Scholar 

  56. Duffy, J. B. GAL4 system in Drosophila: a fly geneticist's Swiss army knife. Genesis 34, 1–15 (2002).

    Article  CAS  PubMed  Google Scholar 

  57. Rorth, P. A modular misexpression screen in Drosophila detecting tissue-specific phenotypes. Proc. Natl Acad. Sci. USA 93, 12418–12422 (1996).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Kooistra, R., Pastink, A., Zonneveld, J. B., Lohman, P. H. & Eeken, J. C. The Drosophila melanogaster DmRAD54 gene plays a crucial role in double-strand break repair after P-element excision and acts synergistically with Ku70 in the repair of X-ray damage. Mol. Cell. Biol. 19, 6269–6275 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Moran, J. V. et al. High frequency retrotransposition in cultured mammalian cells. Cell 87, 917–927 (1996).

    Article  CAS  PubMed  Google Scholar 

  60. Ostertag, E. M. et al. A mouse model of human L1 retrotransposition. Nature Genet. 32, 655–660 (2002). This is the first described model of L1 transposition in the germline of mice.

    Article  CAS  PubMed  Google Scholar 

  61. Han, J. S. & Boeke, J. D. A highly active synthetic mammalian retrotransposon. Nature 429, 314–318 (2004).

    Article  CAS  PubMed  Google Scholar 

  62. Farley, A. H., Luning Prak, E. T. & Kazazian, H. H. Jr. More active human L1 retrotransposons produce longer insertions. Nucleic Acids Res. 32, 502–510 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  63. Gilbert, N., Lutz-Prigge, S. & Moran, J. V. Genomic deletions created upon LINE-1 retrotransposition. Cell 110, 315–325 (2002).

    Article  CAS  PubMed  Google Scholar 

  64. Dewannieux, M., Dupressoir, A., Harper, F., Pierron, G. & Heidmann, T. Identification of autonomous IAP LTR retrotransposons mobile in mammalian cells. Nature Genet. 36, 534–539 (2004).

    Article  CAS  PubMed  Google Scholar 

  65. Esnault, C. et al. APOBEC3G cytidine deaminase inhibits retrotransposition of endogenous retroviruses. Nature 433, 430–433 (2005).

    Article  CAS  PubMed  Google Scholar 

  66. Koga, A., Suzuki, M., Inagaki, H., Bessho, Y. & Hori, H. Transposable element in fish. Nature 383, 30 (1996). This paper reports the identification of the first active endogenous DNA-based transposon in vertebrate species.

    Article  CAS  PubMed  Google Scholar 

  67. Koga, A. et al. The medaka fish Tol2 transposable element can undergo excision in human and mouse cells. J. Hum. Genet. 48, 231–235 (2003).

    Article  CAS  PubMed  Google Scholar 

  68. Kawakami, K. & Noda, T. Transposition of the Tol2 element, an Ac-like element from the Japanese medaka fish Oryzias latipes, in mouse embryonic stem cells. Genetics 166, 895–899 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  69. Klinakis, A. G., Zagoraiou, L., Vassilatis, D. K. & Savakis, C. Genome-wide insertional mutagenesis in human cells by the Drosophila mobile element Minos. EMBO Rep. 1, 416–421 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  70. Zagoraiou, L. et al. In vivo transposition of Minos, a Drosophila mobile element, in mammalian tissues. Proc. Natl Acad. Sci. USA 98, 11474–11478 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  71. Drabek, D. et al. Transposition of the Drosophila hydei Minos transposon in the mouse germ line. Genomics 81, 108–111 (2003).

    Article  CAS  PubMed  Google Scholar 

  72. Schouten, G. J., van Luenen, H. G., Verra, N. C., Valerio, D. & Plasterk, R. H. Transposon Tc1 of the nematode Caenorhabditis elegans jumps in human cells. Nucleic Acids Res. 26, 3013–3017 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. Zhang, L., Sankar, U., Lampe, D. J., Robertson, H. M. & Graham, F. L. The Himar1 mariner transposase cloned in a recombinant adenovirus vector is functional in mammalian cells. Nucleic Acids Res. 26, 3687–3693 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  74. Ivics, Z., Hackett, P. B., Plasterk, R. H. & Izsvak, Z. Molecular reconstruction of Sleeping Beauty, a Tc1-like transposon from fish, and its transposition in human cells. Cell 91, 501–510 (1997). This paper describes the resurrection of the Sleeping Beauty transposon system through site-directed mutagenesis to restore a consensus sequence based on several inactive salmon fish transposase genes.

    Article  CAS  PubMed  Google Scholar 

  75. Luo, G., Ivics, Z., Izsvak, Z. & Bradley, A. Chromosomal transposition of a Tc1/mariner-like element in mouse embryonic stem cells. Proc. Natl Acad. Sci. USA 95, 10769–10773 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  76. Dupuy, A. J. et al. Mammalian germ-line transgenesis by transposition. Proc. Natl Acad. Sci. USA 99, 4495–4499 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  77. Belur, L. R. et al. Gene insertion and long-term expression in lung mediated by the sleeping beauty transposon system. Mol. Ther. 8, 501–507 (2003).

    Article  CAS  PubMed  Google Scholar 

  78. Montini, E. et al. In vivo correction of murine tyrosinemia type I by DNA-mediated transposition. Mol. Ther. 6, 759–769 (2002).

    Article  CAS  PubMed  Google Scholar 

  79. Yant, S. R. et al. Somatic integration and long-term transgene expression in normal and haemophilic mice using a DNA transposon system. Nature Genet. 25, 35–41 (2000).

    Article  CAS  PubMed  Google Scholar 

  80. Ohlfest, J. R., Lobitz, P. D., Perkinson, S. G. & Largaespada, D. A. Integration and long-term expression in xenografted human glioblastoma cells using a plasmid-based transposon system. Mol. Ther. 10, 260–268 (2004).

    Article  CAS  PubMed  Google Scholar 

  81. Dupuy, A. J., Fritz, S. & Largaespada, D. A. Transposition and gene disruption in the male germline of the mouse. Genesis 30, 82–88 (2001).

    Article  CAS  PubMed  Google Scholar 

  82. Fischer, S. E., Wienholds, E. & Plasterk, R. H. Regulated transposition of a fish transposon in the mouse germ line. Proc. Natl Acad. Sci. USA 98, 6759–6764 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  83. Horie, K. et al. Efficient chromosomal transposition of a Tc1/mariner-like transposon Sleeping Beauty in mice. Proc. Natl Acad. Sci. USA 98, 9191–9196 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  84. Carlson, C. M. et al. Transposon mutagenesis of the mouse germline. Genetics 165, 243–256 (2003).

    CAS  PubMed  PubMed Central  Google Scholar 

  85. Horie, K. et al. Characterization of Sleeping Beauty transposition and its application to genetic screening in mice. Mol. Cell. Biol. 23, 9189–9207 (2003). References 81–85 describe Sleeping Beauty -mediated transposition in the germline of mice doubly transgenic for transposon and transposase and its application to mouse genetics and in vivo gene-trapping.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  86. Kile, B. T. et al. Functional genetic analysis of mouse chromosome 11. Nature 425, 81–86 (2003). This paper combined ENU mutagenesis with a balancer chromosome to facilitate a three-generation region-specific screen for mutant phenotypes.

    Article  CAS  PubMed  Google Scholar 

  87. Yant, S. R. et al. High-resolution genome-wide mapping of transposon integration in mammals. Mol. Cell. Biol. 25, 2085–2094 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  88. Wu, X., Li, Y., Crise, B. & Burgess, S. M. Transcription start regions in the human genome are favored targets for MLV integration. Science 300, 1749–1751 (2003).

    Article  CAS  PubMed  Google Scholar 

  89. Jonkers, J. & Berns, A. Retroviral insertional mutagenesis as a strategy to identify cancer genes. Biochim. Biophys. Acta 1287, 29–57 (1996).

    PubMed  Google Scholar 

  90. Carmell, M. A., Zhang, L., Conklin, D. S., Hannon, G. J. & Rosenquist, T. A. Germline transmission of RNAi in mice. Nature Struct Biol 10, 91–92 (2003).

    Article  CAS  PubMed  Google Scholar 

  91. Tiscornia, G., Singer, O., Ikawa, M. & Verma, I. M. A general method for gene knockdown in mice by using lentiviral vectors expressing small interfering RNA. Proc. Natl Acad. Sci. USA 100, 1844–1088 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  92. Berns, K. et al. A large-scale RNAi screen in human cells identifies new components of the p53 pathway. Nature 428, 431–437 (2004).

    Article  CAS  PubMed  Google Scholar 

  93. Paddison, P. J. et al. A resource for large-scale RNA-interference-based screens in mammals. Nature 428, 427–431 (2004).

    Article  CAS  PubMed  Google Scholar 

  94. Beckers, J. & Hrabe de Angelis, M. Large-scale mutational analysis for the annotation of the mouse genome. Curr. Opin. Chem. Biol. 6, 17–23 (2002).

    Article  CAS  PubMed  Google Scholar 

  95. Lewandoski, M. Conditional control of gene expression in the mouse. Nature Rev. Genet. 2, 743–755 (2001).

    Article  CAS  PubMed  Google Scholar 

  96. Townley, D. J., Avery, B. J., Rosen, B. & Skarnes, W. C. Rapid sequence analysis of gene trap integrations to generate a resource of insertional mutations in mice. Genome Res. 7, 293–298 (1997).

    Article  CAS  PubMed  Google Scholar 

  97. Izsvak, Z., Ivics, Z. & Plasterk, R. H. Sleeping Beauty, a wide host-range transposon vector for genetic transformation in vertebrates. J. Mol. Biol. 302, 93–102 (2000).

    Article  CAS  PubMed  Google Scholar 

  98. Yusa, K., Takeda, J. & Horie, K. Enhancement of Sleeping Beauty transposition by CpG methylation: possible role of heterochromatin formation. Mol. Cell. Biol. 24, 4004–4018 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  99. Zayed, H., Izsvak, Z., Khare, D., Heinemann, U. & Ivics, Z. The DNA-bending protein HMGB1 is a cellular cofactor of Sleeping Beauty transposition. Nucleic Acids Res. 31, 2313–2322 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  100. Yant, S. R. & Kay, M. A. Nonhomologous-end-joining factors regulate DNA repair fidelity during Sleeping Beauty element transposition in mammalian cells. Mol. Cell. Biol. 23, 8505–8518 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  101. Izsvak, Z. et al. Healing the wounds inflicted by sleeping beauty transposition by double-strand break repair in mammalian somatic cells. Mol. Cell 13, 279–290 (2004).

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Genetics, Cell Biology, and Development, University of Minnesota Cancer Center, Minneapolis, 55455, Minnesota, USA

    Corey M. Carlson & David A. Largaespada

Authors
  1. Corey M. Carlson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. David A. Largaespada
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to David A. Largaespada.

Ethics declarations

Competing interests

David Largaespada is a co-founder and part owner of the biotechnology company Discovery Genetics Inc (DGI). DGI has licensed Sleeping Beauty technology from the University of Minnesota. DGI is pursuing the use of Sleeping Beauty for human gene therapy.

Related links

Related links

FURTHER INFORMATION

Ensembl

International Gene-trap Consortium

MICER

NCBI

Omnibank

Regeneron Pharmaceuticals

The Arnold and Mabel Beckman Center for Transposon Research

Nature Biotechnology

Glossary

ES CELL LIBRARY

A collection of embryonic stem cell clones that contain a mutation at a distinct location of the genome.

INSERTIONAL MUTAGENESIS

A mutation caused by the addition of DNA to effectively disrupt or alter the function of a given gene.

ALLELIC SERIES

A panel of multiple mutations at distinct locations within the same gene, sometimes resulting in phenotypes that vary in severity or that are completely distinct.

CHROMOSOME ENGINEERING

The controlled generation of chromosomal deletions, inversions or translocations with defined endpoints.

NONSENSE-MEDIATED mRNA DECAY

The process by which the cell destroys mRNAs that are untranslatable owing to the presence of a nonsense codon within the coding region.

EXPRESSION TRAPPING

The identification of ES cell clones from a library that contains gene-trap integrations within genes that are transcriptionally responsive to specific exogenous stimuli.

HAPLOINSUFFICIENCY

The inability for the remaining wild-type allele to compensate for a heterozygous loss-of-function mutation.

PRODRUG METRONIDAZOLE

A compound that when activated by nitrosoreductase is converted into a toxic alkylating agent.

BLOOM SYNDROME

Human autosomal recessive disorder caused by a loss-of-function mutation in a RecQ helicase. It is characterized by genomic instability, increased incidence of sister chromatid exchange, and predisposition to an array of tumour types.

RETROTRANSPOSONS

Transposable elements that mobilize by a 'copy and paste' mechanism. Retrotransposons are transcribed and then reverse transcribed by element-encoded enzymes. The resulting elements are subsequently integrated in the genomes, at a new location (for example, LINE-1 and intracisternal A particle).

DNA TRANSPOSONS

Elements that mobilize by a 'cut and paste' mechanism with the transposon DNA being excised from its original location and inserted elsewhere by the transposase enzyme.

TC1/MARINER TRANSPOSONS

DNA-based 'cut and paste' transposable elements found in numerous host organisms and having a small target sequence for integration (for example, Sleeping Beauty, Himar, Minos).

CONTIGUOUS GENE SYNDROME

A multi-symptom disorder that is caused by a deletion of a large sequence of DNA that encodes several genes.

BALANCER CHROMOSOME

A chromosome with an inversion (or multiple inversions) of an interval of DNA that represses meiotic recombination.

HYPOMORPHIC ALLELE

A mutation in a gene that results in reduced expression or activity of the gene without complete loss of function.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Carlson, C., Largaespada, D. Insertional mutagenesis in mice: new perspectives and tools. Nat Rev Genet 6, 568–580 (2005). https://doi.org/10.1038/nrg1638

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/nrg1638

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing