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Blood, 15 January 2006, Vol. 107, No. 2, pp. 483-491.
Prepublished online as a Blood First Edition Paper on September 27, 2005; DOI 10.1182/blood-2005-05-2133.
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GENE THERAPY
Stable gene transfer and expression in human primary T cells by the Sleeping Beauty transposon system
Xin Huang,
Andrew C. Wilber,
Lei Bao,
Dong Tuong,
Jakub Tolar,
Paul J. Orchard,
Bruce L. Levine,
Carl H. June,
R. Scott McIvor,
Bruce R. Blazar, and
Xianzheng Zhou
From the Division of Blood and Marrow Transplantation, Department of Pediatrics; the Department of Genetics, Cell Biology and Development, University of Minnesota Cancer Center, Minneapolis, MN; and the Department of Pathology and Laboratory Medicine, the Abramson Family Cancer Research Institute, University of Pennsylvania, Philadelphia, PA.
The Sleeping Beauty (SB) transposon system is a nonviral DNA delivery system in which a transposase directs integration of an SB transposon into TA-dinucleotide sites in the genome. To determine whether the SB transposon system can mediate stable gene expression in human T cells, primary peripheral blood lymphocytes (PBLs) were nucleofected with SB vectors carrying a DsRed reporter gene. Plasmids containing the SB transposase on the same molecule as (cis) or on a molecule separate from (trans) the SB transposon mediated long-term and stable reporter gene expression in human primary T cells. Sequencing of transposon:chromosome junctions confirmed that stable gene expression was due to SB-mediated transposition. In other studies, PBLs were successfully transfected using the SB transposon system and shown to stably express a fusion protein consisting of (1) a surface receptor useful for positive T-cell selection and (2) a "suicide" gene useful for elimination of transfected T cells after chemotherapy. This study is the first report demonstrating that the SB transposon system can mediate stable gene transfer in human primary PBLs, which may be advantageous for T-cell–based gene therapies.
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