Abstract
Understanding the functional landscape of the mammalian genome is the next big challenge of biomedical research. The completion of the first phases of the mouse and human genome projects, and expression analyses using microarray hybridization, generate critically important questions about the functional landscape and structure of the mammalian genome: how many genes, and of what type, are there; what kind of functional elements make up a properly functioning gene? One step in this process will be to create mutations in every identifiable mouse gene and analyze the resultant phenotypes. Transposons are being considered as tools to further initiatives to create a comprehensive resource of mutant mouse strains. Also, it may be possible to use transposons in true forward genetic screens in the mouse. The “Sleeping Beauty” (SB) transposon system is one such tool. Moreover, due to its tendency for local hopping, SB has been proposed as a method for regional saturation mutagenesis of the mouse genome. In this chapter, we review the tools and methods currently available to create mutant mice using in vivo, germline transposition in mice.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Ostertag EM, Kazazian HH, Jr. Biology of mammalian L1 retrotransposons. Annu Rev Genet 2001; 35:501–38.
Plasterk RH, Izsvak Z, Ivics Z. Resident aliens: the Tc1/mariner superfamily of transposable elements. Trends Genet 1999; 15:326–32.
Ivics Z, Hackett PB, Plasterk RH, Izsvak Z. Molecular reconstruction of Sleeping Beauty, a Tc1-like transposon from fish, and its transposition in human cells. Cell 1997; 91:501–10.
Cui Z, Geurts AM, Liu G, Kaufman CD, Hackett PB. Structure–function analysis of the inverted terminal repeats of the Sleeping Beauty transposon. J Mol Biol 2002; 318:1221–35.
Geurts AM, Yang Y, Clark KJ, et al. Gene transfer into genomes of human cells by the Sleeping Beauty transposon system. Mol Ther 2003; 8:108–17.
Yant SR, Park J, Huang Y, Mikkelsen JG, Kay MA. Mutational analysis of the N-terminal DNA-binding domain of Sleeping Beauty transposase: critical residues for DNA binding and hyperactivity in mammalian cells. Mol Cell Biol 2004; 24:9239–47.
Zayed H, Izsvak Z, Walisko O, Ivics Z. Development of hyperactive Sleeping Beauty transposon vectors by mutational analysis. Mol Ther 2004; 9:292–304.
Miskey C, Izsvak Z, Kawakami K, Ivics Z. DNA transposons in vertebrate functional genomics. Cell Mol Life Sci 2005; 62:629–41.
Collier LS, Carlson CM, Ravimohan S, Dupuy AJ, Largaespada DA. Cancer gene discovery in solid tumours using transposon-based somatic mutagenesis in the mouse. Nature 2005; 436:272–6.
Dupuy AJ, Akagi K, Largaespada DA, Copeland NG, Jenkins NA. Mammalian mutagenesis using a highly mobile somatic Sleeping Beauty transposon system. Nature 2005; 436:221–6.
Drabek D, Zagoraiou L, deWit T, et al. Transposition of the Drosophila hydei Minos transposon in the mouse germ line. Genomics 2003; 81:108–11.
Ding S, Wu X, Li G, Han M, Zhuang Y, Xu T. Efficient transposition of the piggyBac (PB) transposon in mammalian cells and mice. Cell 2005; 122:473–83.
Wu S, Ying G, Wu Q, Capecchi MR. Toward simpler and faster genome-wide mutagenesis in mice. Nat Genet 2007; 39:922–30.
Dupuy AJ, Fritz S, Largaespada DA. Transposition and gene disruption in the male germline of the mouse. Genesis 2001; 30:82–8.
Roberg-Perez K, Carlson CM, Largaespada DA. MTID: a database of Sleeping Beauty transposon insertions in mice. Nucleic Acids Res 2003; 31:78–81.
Carlson CM, Dupuy AJ, Fritz S, Roberg-Perez KJ, Fletcher CF, Largaespada DA. Transposon mutagenesis of the mouse germline. Genetics 2003; 165:243–56.
Geurts AM, Collier LS, Geurts JL, et al. Gene mutations and genomic rearrangements in the mouse as a result of transposon mobilization from chromosomal concatemers. PLoS Genet 2006; 2:e156.
Geurts AM, Wilber A, Carlson CM, et al. Conditional gene expression in the mouse using a Sleeping Beauty gene-trap transposon. BMC Biotechnol 2006; 6:30.
Keng VW, Yae K, Hayakawa T, et al. Region-specific saturation germline mutagenesis in mice using the Sleeping Beauty transposon system. Nat Methods 2005; 2:763–9.
Fischer SE, Wienholds E, Plasterk RH. Regulated transposition of a fish transposon in the mouse germ line. Proc Natl Acad Sci USA 2001; 98:6759–64.
Horie K, Kuroiwa A, Ikawa M, et al. Efficient chromosomal transposition of a Tc1/mariner-like transposon Sleeping Beauty in mice. Proc Natl Acad Sci USA 2001; 98:9191–6.
Horie K, Yusa K, Yae K, et al. Characterization of Sleeping Beauty transposition and its application to genetic screening in mice. Mol Cell Biol 2003; 23:9189–207.
Kitada K, Ishishita S, Tosaka K, et al. Transposon-tagged mutagenesis in the rat. Nat Methods 2007; 4:131–3.
Lu B, Geurts AM, Poirier C, et al. Generation of rat mutants using a coat color-tagged Sleeping Beauty transposon system. Mamm Genome2007; 18:338–46.
Ryder E, Russell S. Transposable elements as tools for genomics and genetics in Drosophila. Brief Funct. Genomic Proteomic 2003; 2:57–71.
Stanford WL, Cohn JB, Cordes SP. Gene-trap mutagenesis: past, present and beyond. Nat Rev Genet 2001; 2:756–68.
Abuin A, Hansen GM, Zambrowicz B. Gene trap mutagenesis. Handb Exp Pharmacol 2007: 129–47.
Yusa K, Takeda J, Horie K. Enhancement of Sleeping Beauty transposition by CpG methylation: possible role of heterochromatin formation. Mol Cell Biol 2004; 24:4004–18.
Vigdal TJ, Kaufman CD, Izsvak Z, Voytas DF, Ivics Z. Common physical properties of DNA affecting target site selection of Sleeping Beauty and other Tc1/mariner transposable elements. J Mol Biol 2002; 323:441–52.
Okabe M, Ikawa M, Kominami K, Nakanishi T, Nishimune Y. “Green mice” as a source of ubiquitous green cells. FEBS Lett 1997; 407:313–9.
Okabe M, Ikawa M, Kominami K, Nakanishi T, Nishimune Y. “Green mice” as a source of ubiquitous green cells. FEBS Lett 1997; 407:313–9.
Dupuy AJ, Akagi K, Largaespada DA, Copeland NG, Jenkins NA. Mammalian mutagenesis using a highly mobile somatic Sleeping Beauty transposon system. Nature 2005; 436:221–6.
Roths JB, Foxworth WB, McArthur MJ, Montgomery CA, Kier AB. Spontaneous and engineered mutant mice as models for experimental and comparative pathology: history, comparison, and developmental technology. Lab Anim Sci 1999; 49:12–34.
Geurts AM, Yang Y, Clark KJ, et al. Gene transfer into genomes of human cells by the Sleeping Beauty transposon system. Mol Ther 2003; 8:108–17.
Acknowledgments
This work was supported by grants R21 CA118600, R01 CA113636, and RO1 DA014764 from the National Institutes of Health (to DAL).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Largaespada, D.A. (2009). Transposon Mutagenesis in Mice. In: Wurst, W., Kühn, R. (eds) Gene Knockout Protocols. Methods in Molecular Biology, vol 530. Humana Press. https://doi.org/10.1007/978-1-59745-471-1_20
Download citation
DOI: https://doi.org/10.1007/978-1-59745-471-1_20
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-934115-26-8
Online ISBN: 978-1-59745-471-1
eBook Packages: Springer Protocols