title: Stable gene transfer and expression in human primary T cells by the Sleeping Beauty transposon system.
abstract: 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.
authors.affiliation: Division of Blood and Marrow Transplantation, Department of Pediatrics, University of Minnesota Cancer Center, Minneapolis, MN 55455,USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Huang
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Wilber
authors.forename: Lei
authors.initials: L
authors.lastname: Bao
authors.forename: Dong
authors.initials: D
authors.lastname: Tuong
authors.forename: Jakub
authors.initials: J
authors.lastname: Tolar
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Orchard
authors.forename: Bruce L
authors.initials: BL
authors.lastname: Levine
authors.forename: Carl H
authors.initials: CH
authors.lastname: June
authors.forename: R Scott
authors.initials: RS
authors.lastname: McIvor
authors.forename: Bruce R
authors.initials: BR
authors.lastname: Blazar
authors.forename: Xianzheng
authors.initials: X
authors.lastname: Zhou
mesh_terms: D000818:Animals; D004251:DNA Transposable Elements / Q000235:genetics*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000502:physiology; D005822:Genetic Vectors; D006801:Humans; D008213:Lymphocyte Activation; D010957:Plasmids / Q000494:pharmacology*; D013552:Swine; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D014162:Transfection; D019076:Transgenes / Q000502:physiology*; D019895:Transposases / Q000235:genetics* / Q000378:metabolism*
chemical_list: D004251:DNA Transposable Elements; D019895:Transposases; C494471:sleeping beauty transposase, human