Skip to main content

Insertional Mutagenesis of the Mouse Germline With Sleeping Beauty Transposition

  • Protocol
Book cover Chromosomal Mutagenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 435))

Abstract

Efficient linking of primary DNA sequence information to gene functions in vertebrate models requires that genetic modifications and their effects are analyzed in an efficacious, controlled, and scalable manner. Thus, to facilitate analysis of gene function, new genetic tools and strategies are currently under development. Transposable elements, by virtue of their inherent ability to insert into DNA, can be developed into useful tools for chromosomal manipulations. Mutagenesis screens based on transposable elements have numerous advantages as they can be applied in vivo and are therefore phenotype-driven, and molecular analysis of the mutations is straightforward. Current progress in this field indicates that transposable elements will serve as indispensable tools in the genetic toolkit of vertebrate models. Here, we provide experimental protocols for the construction, functional testing, and application of the Sleeping Beauty transposon for insertional mutagenesis of the mouse germline.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Ivics, Z., Hackett, P. B., Plasterk, R. H., 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.

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  3. Luo, G., Ivics, Z., Izsvak, Z., and Bradley, A. (1998) Chromosomal transposition of a Tc1/mariner-like element in mouse embryonic stem cells. Proc. Natl. Acad. Sci. USA 95, 10,769–10,773.

    Article  CAS  PubMed  Google Scholar 

  4. Yant, S. R., Meuse, L., Chiu, W., Ivics, Z., Izsvak, Z., and Kay, M. A. (2000) Somatic integration and long-term transgene expression in normal and haemophilic mice using a DNA transposon system. Nat. Genet. 25, 35–41.

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  7. Horie, K., Kuroiwa, A., Ikawa, M., Okabe, M., Kondoh, G., Matsuda, Y., and Takeda, J. (2001) Efficient chromosomal transposition of a Tc1/mariner-like transposon Sleeping Beauty in mice. Proc. Natl. Acad. Sci. USA 98, 9191–9196.

    Article  CAS  PubMed  Google Scholar 

  8. Horie, K., Yusa, K., Yae, K., et al. (2003) Characterization of Sleeping Beauty transposition and its application to genetic screening in mice. Mol. Cell. Biol. 23, 9189–9207.

    Article  CAS  PubMed  Google Scholar 

  9. Carlson, C. M., Dupuy, A. J., Fritz, S., Roberg-Perez, K. J., Fletcher, C. F., and Largaespada, D. A. (2003) Transposon mutagenesis of the mouse germline. Genetics 165, 243–256.

    CAS  PubMed  Google Scholar 

  10. Kitada, K., Ishishita, S., Tosaka, K., et al. (2007) Transposon-tagged mutagenesis in the rat. Nat. Methods 4, 131–133.

    Article  CAS  PubMed  Google Scholar 

  11. Miskey, C., Izsvak, Z., Kawakami, K., and Ivics, Z. (2005) DNA transposons in vertebrate functional genomics. Cell. Mol. Life Sci. 62, 629–641.

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  13. Friedrich, G. and Soriano, P. (1991) Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice. Genes Dev. 5, 1513–1523.

    Article  CAS  PubMed  Google Scholar 

  14. Zambrowicz, B. P., Friedrich, G. A., Buxton, E. C., Lilleberg, S. L., Person, C., and Sands, A. T. (1998) Disruption and sequence identification of 2,000 genes in mouse embryonic stem cells. Nature 392, 608–611.

    Article  CAS  PubMed  Google Scholar 

  15. Clark, K. J., Geurts, A. M., Bell, J. B., and Hackett, P. B. (2004) Transposon vectors for gene-trap insertional mutagenesis in vertebrates. Genesis 39, 225–233.

    Article  CAS  PubMed  Google Scholar 

  16. Geurts, A. M., Wilber, A., Carlson, C. M., et al. (2006) Conditional gene expression in the mouse using a Sleeping Beauty gene-trap transposon. BMC Biotechnol. 6, 30.

    Article  PubMed  Google Scholar 

  17. Yae, K., Keng, V. W., Koike, M., et al. (2006) Sleeping beauty transposon-based phenotypic analysis of mice: lack of Arpc3 results in defective trophoblast outgrowth. Mol. Cell. Biol. 26, 6185–6196.

    Article  CAS  PubMed  Google Scholar 

  18. Keng, V. W., Yae, K., Hayakawa, T., et al. (2005) Region-specific saturation germline mutagenesis in mice using the Sleeping Beauty transposon system. Nat. Methods. 2, 763–769.

    Article  CAS  PubMed  Google Scholar 

  19. Ishida, Y. and Leder, P. (1999) RET: a poly A-trap retrovirus vector for reversible disruption and expression monitoring of genes in living cells. Nucleic Acids Res. 27, E35

    Article  CAS  PubMed  Google Scholar 

  20. Baus, J., Liu, L., Heggestad, A. D., Sanz, S., and Fletcher, B. S. (2005) Hyperactive transposase mutants of the Sleeping Beauty transposon. Mol. Ther. 12, 1148–1156.

    Article  CAS  PubMed  Google Scholar 

  21. Geurts, A. M., Yang, Y., Clark, K. J., et al. (2003) Gene transfer into genomes of human cells by the sleeping beauty transposon system. Mol. Ther. 8, 108–117.

    Article  CAS  PubMed  Google Scholar 

  22. Zayed, H., Izsvak, Z., Walisko, O., and Ivics, Z. (2004) Development of hyperactive sleeping beauty transposon vectors by mutational analysis. Mol. Ther. 9, 292–304.

    Article  CAS  PubMed  Google Scholar 

  23. Yant, S. R., Park, J., Huang, Y., Mikkelsen, J. G., and Kay, M. A. (2004) 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. 24, 9239–9247.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2008 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Takeda, J., Izsvák, Z., Ivics, Z. (2008). Insertional Mutagenesis of the Mouse Germline With Sleeping Beauty Transposition. In: Davis, G.D., Kayser, K.J. (eds) Chromosomal Mutagenesis. Methods in Molecular Biology, vol 435. Humana Press. https://doi.org/10.1007/978-1-59745-232-8_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-232-8_8

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-899-7

  • Online ISBN: 978-1-59745-232-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics