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, Xianzheng Zhou

Research output: Contribution to journalArticlepeer-review

90 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)483-491
Number of pages9
JournalBlood
Volume107
Issue number2
DOIs
StatePublished - Jan 15 2006

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