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Stable gene transfer and expression in human primary T cells by the Sleeping Beauty transposon system.

Authors
  • Huang, Xin
  • Wilber, Andrew C
  • Bao, Lei
  • Tuong, Dong
  • Tolar, Jakub
  • Orchard, Paul J
  • Levine, Bruce L
  • June, Carl H
  • McIvor, R Scott
  • Blazar, Bruce R
  • Zhou, Xianzheng
Type
Published Article
Journal
Blood
Publication Date
Jan 15, 2006
Volume
107
Issue
2
Pages
483–491
Identifiers
PMID: 16189271
Source
Medline
License
Unknown

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.

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