Skip to main content
SpringerLink
Log in
Book cover

Gene Therapy Protocols pp 267–276Cite as

Liver-Directed Gene Therapy Using the Sleeping Beauty Transposon System

  • Protocol

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 434))

Summary

Sleeping Beauty (SB) is a transposon system genetically reconstructed from teleost fish that mediates chromosomal integration of DNA sequences by a cut-and-paste mechanism. SB has been shown to mediate transposition in a variety of cells and tissues, has been used for the generation of transgenic animals and has been tested as a vector for gene therapy in several animal models of human disease. Here, we describe methods that we have developed for testing SB-mediated transposition, first in cultured mammalian cells, and then in vivo using a combination of rapid, high-volume tail vein injection for DNA delivery to the liver along with in vivo bioluminescence imaging to monitor sustained luciferase gene expression in individual animals.

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

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
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

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

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. Geurts, A. M., Hackett, C. S., Bell, J. B., Bergemann, T. L., Collier, L. S., Carlson, C. M., Largaespada, D. A., and Hackett, P. B. (2006) Structure-based prediction of insertion-site preferences of transposons into chromosomes. Nucleic Acids Research 34, 2803–2811.

    Article  CAS  PubMed  Google Scholar 

  3. 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 

  4. Yant, S. R., Wu, X., Huang, Y., Garrison, B., Burgess, S. M., and Kay, M. A. (2005) High-resolution genome-wide mapping of transposon integration in mammals Molecular Cellular Biology 25, 2085–2094.

    Article  CAS  Google Scholar 

  5. Davidson, A. E., Balciunas, D., Mohn, D., Shaffer, J., Hermanson, S., Sivasubbu, S., Cliff, M. P., Hackett, P. B., and Ekker, S. C. (2003) Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon. Developmental Biology 263, 191–202.

    Article  CAS  PubMed  Google Scholar 

  6. Clark, K. J., Geurts, A. M., Bell, J. B., and Hackett, P. B. (2004) Transposon vectors for gene-trap insertional mutagenesis in vertebrates. Genesis: the Journal of Genetics Development 39, 225–233.

    CAS  Google Scholar 

  7. Wadman, S. A., Clark, K. J., and Hackett, P. B. (2005) Fishing for answers with transposons. Marine Biotechnology 7, 135–141.

    Article  CAS  PubMed  Google Scholar 

  8. Dupuy, A. J., Clark, K., Carlson, C. M., Fritz, S., Davidson, A. E., Markley, K. M., Finley, K., Fletcher, C. F., Ekker, S. C., Hackett, P. B., Horn, S., and Largaespada, D. A. (2002) Mammalian germ-line transgenesis by transposition. Proceedings of the National Academy of Sciences of the United States of America 99, 44954–44959.

    Article  Google Scholar 

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

    CAS  Google Scholar 

  10. Fischer, S. E., Wienholds, E., and Plasterk, R. H. (2001) Regulated transposition of a fish transposon in the mouse germ line. Proceedings of the National Academy of Sciences of the United States of America 98, 6759–6764.

    Article  CAS  PubMed  Google Scholar 

  11. 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. Proceedings of the National Academy of Sciences of the United States of America 98, 9191–9196.

    Article  CAS  PubMed  Google Scholar 

  12. Horie, K., Yusa, K., Yae, K., Odajima, J., Fischer, S. E., Keng, V. W., Hayakawa, T., Mizuno, S., Kondoh, G., Ijiri, T., Matsuda, Y., Plasterk, R. H., and Takeda, J. (2003) Characterization of Sleeping Beauty transposition and its application to genetic screening in mice. Molecular Cellular Biology 23, 9189–9207.

    Article  CAS  Google Scholar 

  13. Huang, X., Wilber, A. C., Bao, L., Tuong, D., Tolar, J., Orchard, P. J., Levine, B. L., June, C. H., McIvor, R. S., Blazar, B. R., and Zhou, X. (2006) Stable gene transfer and expression in human primary T cells by the Sleeping Beauty transposon system. Blood 107, 483–491.

    Article  CAS  PubMed  Google Scholar 

  14. Belur, L. R., Frandsen, J. L., Dupuy, A. J., Ingbar, D. H., Largaespada, D. A., Hackett, P. B., and McIvor, R.S. (2003) Gene insertion and long-term expression in lung mediated by the Sleeping Beauty transposon system. Molecular Therapy: the Journal of the American Society of Gene Therapy 8, 501–507.

    CAS  Google Scholar 

  15. Liu, L., Sanz, S., Heggestad, A. D., Antharam, V., Notterpek, L., and Fletcher, B. S. (2004) Endothelial targeting of the Sleeping Beauty transposon within lung. Molecular Therapy: the Journal of the American Society of Gene Therapy 10, 97–105.

    CAS  Google Scholar 

  16. 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. Nature Genetics 25, 35–41.

    Article  CAS  PubMed  Google Scholar 

  17. Yant, S. R., Ehrhardt, A., Mikkelsen, J. G., Meuse, L., Pham, T., and Kay, M. A. (2002) Transposition from a gutless adeno-transposon vector stabilizes transgene expression in vivo. Nature Biotechnology 20, 999–1005.

    Article  CAS  PubMed  Google Scholar 

  18. Score, P. R., Belur, L. R., Frandsen, J. L., Guerts, J. L., Yamaguchi, T., Somia, N. V., Hackett, P. B., Largaespada, D. A., and McIvor, R. S. (2006) Sleeping Beauty -mediated transposition and long-term expression in vivo: use of the LoxP/Cre recombinase system to distinguish transposition-specific expression. Molecular Therapy 13, 617–624.

    Article  CAS  PubMed  Google Scholar 

  19. Wilber, A., Frandsen, J. F., Geurts, J. L., Largaespada, D. A., Hackett, P. B., and McIvor, R. S. (2006) RNA as a source of transposase for Sleeping Beauty-mediated gene insertion and expression in somatic cells and tissues. Molecular Therapy 13 , 625–630.

    Article  CAS  PubMed  Google Scholar 

  20. Ohlfest, J. R., Frandsen, J. L., Fritz, S., Lobitz, P. D., Perkinson, S. G., Clark, K. J., Nelsestuen, G., Key, N. S., McIvor, R. S., Hackett, P. B., and Largaespada, D. A. Phenotypic correction and long-term expression of factor VIII in hemophilic mice by immunotolerization and nonviral gene transfer using the Sleeping Beauty transposon system. (2005) Blood 105, 2691–2698.

    Article  CAS  PubMed  Google Scholar 

  21. Ohlfest, J. R., Demorest, Z. L., Motooka, Y., Vengco, I., Oh, S., Chen, E., Scappaticci, F. A., Saplis, R. J., Ekker, S. C., Low, W. C., Freese, A. B., and Largaespada, D. A. (2005) Combinatorial antiangiogenic gene therapy by nonviral gene transfer using the Sleeping Beauty transposon causes tumor regression and improves survival in mice bearing intracranial human glioblastoma. Molecular Therapy: the Journal of the American Society of Gene Therapy 12, 778–788.

    CAS  Google Scholar 

  22. Liu, F., Song, Y., and Liu, D. Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. (1999) Gene Therapy 6, 1258–1266.

    Article  CAS  PubMed  Google Scholar 

  23. Zhang, G., Budker, V., and Wolff, J. A. High levels of foreign gene expression in hepatocytes after tail vein injections of naked plasmid DNA. (1999) Hum Gene Ther 10, 1735–1737.

    Article  CAS  PubMed  Google Scholar 

  24. Zhang, G., Gao, X., Song, Y. K., Vollmer, R., Stolz, D. B., Gasiorowski, J. Z., Dean, D. A., and Liu, D. (2004) Hydroporation as the mechanism of hydrodynamic delivery. Gene Therapy 11, 675–682.

    Article  CAS  PubMed  Google Scholar 

  25. Suda, T., Gao, X., Stolz, D. B., and Liu, D. (2007) Structural impact of hydrodynamic injection on mouse liver. Gene Therapy 14, 129–137.

    CAS  PubMed  Google Scholar 

  26. Geurts, A. M., Yang, Y., Clark, K. J., Liu, G., Cui, Z., Dupuy, A. J., Bell, J. B., Largaespada, D. A., and Hackett, P. B. (2003) Gene transfer into genomes of human cells by the Sleeping Beauty transposon system. Molecular Therapy: the Journal of the American Society of Gene Therapy 8, 108–117.

    CAS  Google Scholar 

  27. Mikkelsen, J. G., Yant, S. R., Meuse, L., Huang, Z., Xu, H., and Kay, M. A. (2003) Helper-Independent Sleeping Beauty transposon-transposase vectors for efficient nonviral gene delivery and persistent gene expression in vivo. Molecular Therapy: the Journal of the American Society of Gene Therapy 8, 654–665.

    CAS  Google Scholar 

  28. Wilber, A., Frandsen, J. L., Wangensteen, K. J., Ekker, S. C., Wang, X., and McIvor, R. S. (2005) Dynamic gene expression after systemic delivery of plasmid DNA as determined by in vivo bioluminescence imaging. Human Gene Therapy 16,1325–1332.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Arnold and Mabel Beckman Center for Transposon Research, Gene Therapy Program, Institute of Human Genetics, and Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN

    Lalitha R. Belur, R. Scott McIvor & Andrew Wilber

Authors
  1. Lalitha R. Belur
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R. Scott McIvor
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Andrew Wilber
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30322-0355

    Joseph M. Le Doux

Rights and permissions

Reprints and permissions

Copyright information

© 2008 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Belur, L.R., McIvor, R.S., Wilber, A. (2008). Liver-Directed Gene Therapy Using the Sleeping Beauty Transposon System. In: Le Doux, J.M. (eds) Gene Therapy Protocols. Methods in Molecular Biology™, vol 434. Humana Press. https://doi.org/10.1007/978-1-60327-248-3_16

Download citation

  • DOI: https://doi.org/10.1007/978-1-60327-248-3_16

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60327-247-6

  • Online ISBN: 978-1-60327-248-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation