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Preferential delivery of the Sleeping Beauty transposon system to livers of mice by hydrodynamic injection

Nature Protocols volume 2, pages 31533165 (2007) | Download Citation

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Abstract

Nonviral, DNA-mediated gene transfer is an alternative to viral delivery systems for expressing new genes in cells and tissues. The Sleeping Beauty (SB) transposon system combines the advantages of viruses and naked DNA molecules for gene therapy purposes; however, efficacious delivery of DNA molecules to animal tissues can still be problematic. Here we describe the hydrodynamic delivery procedure for the SB transposon system that allows efficient delivery to the liver in the mouse. The procedure involves rapid, high-pressure injection of a DNA solution into the tail vein. The overall procedure takes <1 h although the delivery into one mouse requires only a few seconds. Successful injections result in expression of the transgene in 5–40% of hepatocytes 1 d after injection. Several weeks after injection, transgene expression stabilizes at 1% of the level at 24 h, presumably owing to integration of the transposons into chromosomes.

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Acknowledgements

We thank the Arnold and Mabel Beckman Foundation for support of our work and all members of the Beckman Center for Transposon Research for a long history of contributions of ideas and results. We appreciate the help of Mr. Joel Frandsen for teaching us the intricacies of hydrodynamic injection and our veterinarian technician, Brenda Koniar. The authors were partially supported by National Institutes of Health grant 1PO1 HD32652-07.

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Affiliations

  1. Department of Genetics, Cell Biology and Development, Beckman Center for Transposon Research, Institute of Human Genetics, University of Minnesota, 6-160 Jackson Hall, 321 Church Street SE, Minneapolis, Minnesota 55455, USA.

    • Jason B Bell
    • , Kelly M Podetz-Pedersen
    • , Elena L Aronovich
    • , Lalitha R Belur
    • , R Scott McIvor
    •  & Perry B Hackett
  2. Discovery Genomics, Inc., 614 McKinley Place NE, Minneapolis, Minnesota 55413, USA.

    • R Scott McIvor

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Competing interests

R.S.M. and P.B.H. declare a competing financial interest; the other authors declare that they have no competing financial interests.

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Correspondence to Perry B Hackett.

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https://doi.org/10.1038/nprot.2007.471

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