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Blood, 1 April 2005, Vol. 105, No. 7, pp. 2691-2698. Prepublished online as a Blood First Edition Paper on December 2, 2004; DOI 10.1182/blood-2004-09-3496. This Article Full Text Full Text (PDF) All Versions of this Article: 2004-09-3496v1 105/7/2691    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Ohlfest, J. R. Articles by Largaespada, D. A. Search for Related Content PubMed PubMed Citation Articles by Ohlfest, J. R. Articles by Largaespada, D. A. Related Collections Hemostasis, Thrombosis, and Vascular Biology Immunotherapy Gene Therapy Related Article in Blood Online Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  GENE THERAPY Phenotypic correction and long-term expression of factor VIII in hemophilic mice by immunotolerization and nonviral gene transfer using the Sleeping Beauty transposon system John R. Ohlfest, Joel L. Frandsen, Sabine Fritz, Paul D. Lobitz, Scott G. Perkinson, Karl J. Clark, Gary Nelsestuen, Nigel S. Key, R. Scott McIvor, Perry B. Hackett, and David A. Largaespada From the Department of Genetics, Cell Biology and Development, University of Minnesota Cancer Center, Arnold and Mabel Beckman Center for Transposon Research, University of Minnesota; Discovery Genomics Inc. Minneapolis, MN; the Department of Biochemistry, Molecular Biology, and Biophysics, University or Minnesota, St Paul, MN; and the Department of Medicine, University of Minnesota, Minneapolis, MN. Hemophilia A is a lead candidate for treatment by gene therapy because small increments in the missing secreted protein product, coagulation factor VIII (FVIII), would result in substantial clinical amelioration. Clinically relevant therapy might be achieved by stably delivering a human FVIII cDNA to correct the bleeding disorder. We used the Sleeping Beauty (SB) transposon, delivered as naked plasmid DNA by tail-vein injection, to integrate B-domain–deleted FVIII genes into the chromosomes of hemophilia A mice and correct the phenotype. Since FVIII protein is a neoantigen to these mice, sustaining therapeutic plasma FVIII levels was problematic due to inhibitory antibody production. We circumvented this problem by tolerizing 82% of neonates by a single facial-vein injection of recombinant FVIII within 24 hours of birth (the remaining 18% formed inhibitors). Achievement of high-level (10%-100% of normal) FVIII expression and phenotypic correction required co-injection of an SB transposase-expressing plasmid to facilitate transgene integration in immunotolerized animals. Linker-mediated polymerase chain reaction was used to clone FVIII transposon insertion sites from liver genomic DNA, providing molecular evidence of transposition. Thus, SB provides a nonviral means for sustained FVIII gene delivery in a mouse model of hemophilia A if the immune response is prevented. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? Related Article in Blood Online: An ounce of prevention potentiates a pound of cure for hemophilia A Katherine Parker Ponder Blood 2005 105: 2620. [Full Text] [PDF] This article has been cited by other articles: T. VandenDriessche, Z. Ivics, Z. Izsvak, and M. K. L. Chuah Emerging potential of transposons for gene therapy and generation of induced pluripotent stem cells Blood, August 20, 2009; 114(8): 1461 - 1468. [Abstract] [Full Text] [PDF] K. J. Clark, D. F. Carlson, M. J. Leaver, L. K. Foster, and S. C. Fahrenkrug Passport, a native Tc1 transposon from flatfish, is functionally active in vertebrate cells Nucleic Acids Res., March 1, 2009; 37(4): 1239 - 1247. [Abstract] [Full Text] [PDF] S. M. Wiesner, S. A. Decker, J. D. Larson, K. Ericson, C. Forster, J. L. Gallardo, C. Long, Z. L. Demorest, E. A. Zamora, W. C. Low, et al. De novo Induction of Genetically Engineered Brain Tumors in Mice Using Plasmid DNA Cancer Res., January 15, 2009; 69(2): 431 - 439. [Abstract] [Full Text] [PDF] L. Liu, H. Liu, G. Visner, and B. S. Fletcher Sleeping Beauty-mediated eNOS gene therapy attenuates monocrotaline-induced pulmonary hypertension in rats FASEB J, December 1, 2006; 20(14): 2594 - 2596. [Abstract] [Full Text] [PDF] H. Liu, L. Liu, B. S. Fletcher, and G. A. Visner Sleeping Beauty-based gene therapy with indoleamine 2,3-dioxygenase inhibits lung allograft fibrosis FASEB J, November 1, 2006; 20(13): 2384 - 2386. [Abstract] [Full Text] [PDF]

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