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Blood, 13 August 2009, Vol. 114, No. 7, pp. 1319-1330. Prepublished online as a Blood First Edition Paper on May 4, 2009; DOI 10.1182/blood-2009-03-210005. This Article Full Text Full Text (PDF) Supplemental Figures All Versions of this Article: blood-2009-03-210005v1 114/7/1319    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 Google Scholar Articles by Xue, X. Articles by Zhou, X. PubMed PubMed Citation Articles by Xue, X. Articles by Zhou, X. Related Collections Gene Therapy Related Article in Blood Online Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  GENE THERAPY Stable gene transfer and expression in cord blood–derived CD34+ hematopoietic stem and progenitor cells by a hyperactive Sleeping Beauty transposon system Xingkui Xue14, Xin Huang14, Sonja E. Nodland2,3,5, Lajos Mátés6, Linan Ma7, Zsuzsanna Izsvák6, Zoltán Ivics6, Tucker W. LeBien2,3,5, R. Scott McIvor2,4,8, John E. Wagner1,2, and Xianzheng Zhou14 1 Division of Pediatric Blood and Marrow Transplantation, 2 Masonic Cancer Center, 3 Center for Immunology, 4 Center for Genome Engineering, 5 Department of Laboratory Medicine and Pathology, and 6 Max Delbrück Center for Molecular Medicine, Berlin, Germany; and 7 Biostatistics and Informatics Core at the Masonic Cancer Center and 8 Department of Genetics, Cell Biology and Development, University of Minnesota Medical School, Minneapolis Here we report stable gene transfer in cord blood-derived CD34+ hematopoietic stem cells using a hyperactive nonviral Sleeping Beauty (SB) transposase (SB100X). In colony-forming assays, SB100X mediated the highest efficiency (24%) of stable Discosoma sp red fluorescent protein (DsRed) reporter gene transfer in committed hematopoietic progenitors compared with both the early-generation hyperactive SB11 transposase and the piggyBac transposon system (1.23% and 3.8%, respectively). In vitro differentiation assays further demonstrated that SB100X-transfected CD34+ cells can develop into DsRed+ CD4+CD8+ T (3.17%-21.84%; median, 7.97%), CD19+ B (3.83%-18.66%; median, 7.84%), CD56+CD3– NK (3.53%-79.98%; median, 7.88%), and CD33+ myeloid (7.59%-15.63%; median, 9.48%) cells. SB100X-transfected CD34+ cells achieved approximately 46% engraftment in NOD-scid IL2cnull (NOG) mice. Twelve weeks after transplantation, 0.57% to 28.96% (median, 2.79%) and 0.49% to 34.50% (median, 5.59%) of total human CD45+ cells in the bone marrow and spleen expressed DsRed, including CD19+ B, CD14+ monocytoid, and CD33+ myeloid cell lineages. Integration site analysis revealed SB transposon sequences in the human chromosomes of in vitro differentiated T, B, NK, and myeloid cells, as well as in human CD45+ cells isolated from bone marrow and spleen of transplanted NOG mice. Our results support the continuing development of SB-based gene transfer into human hematopoietic stem cells as a modality for gene therapy. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? Related Article in Blood Online: Better positioned in stem cells Linzhao Cheng Blood 2009 114: 1285-1286. [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] L. Cheng Better positioned in stem cells Blood, August 13, 2009; 114(7): 1285 - 1286. [Full Text] [PDF]

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