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Stable gene transfer and expression in cord blood-derived CD34+ hematopoietic stem and progenitor cells by a hyperactive Sleeping Beauty transposon system

Official URL:https://doi.org/10.1182/blood-2009-03-210005
PubMed:View item in PubMed
Creators Name:Xue, X. and Huang, X. and Nodland, S.E. and Mates, L. and Ma, L. and Izsvak, Z. and Ivics, Z. and Lebien, T.W. and McIvor, R.S. and Wagner, J.E. and Zhou, X.
Journal Title:Blood
Journal Abbreviation:Blood
Page Range:1319-1330
Date:13 August 2009
Keywords:CD34 Antigens, Differentiation Antigens, Cell Differentiation, Cord Blood Stem Cell Transplantation, DNA Transposable Elements, Fetal Blood, Gene Expression, Gene Therapy, Gene Transfer Techniques, Graft Survival, Hematopoietic Stem Cells, Luminescent Proteins, Time Factors, Heterologous Transplantation, Animals, Mice
Abstract:Here we report stable gene transfer in cord blood-derived CD34(+) hematopoietic stem cells (HSCs) using a hyperactive non-viral 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 to 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 ~46% engraftment in NOD-scid IL2gammac(null) (NOG) mice. Twelve weeks after transplantation, 0.57-28.96% (median=2.79%) and 0.49-34.50% (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 HSCs as a modality for gene therapy.
Publisher:American Society of Hematology (U.S.A.)
Item Type:Article

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