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
Division of Blood and Marrow Transplantation, Department of Pediatrics, University of Minnesota Cancer Center, Minneapolis, MN 55455,USA.
Blood. 2006 Jan 15;107(2):483-91. doi: 10.1182/blood-2005-05-2133. Epub 2005 Sep 27.
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.
睡美人（SB）转座子系统是一种非病毒DNA递送系统，其中转座酶指导SB转座子整合到基因组中的TA二核苷酸位点。为了确定SB转座子系统是否能介导人T细胞中的稳定基因表达，用携带DsRed报告基因的SB载体对原代外周血淋巴细胞（PBL）进行了核转染。与SB转座子在同一分子上（顺式）或在与SB转座子分开的分子上（反式）含有SB转座酶的质粒介导了人原代T细胞中的长期稳定报告基因表达。转座子与染色体连接点的测序证实，稳定的基因表达是由于SB介导的转座。在其他研究中，使用SB转座子系统成功转染了PBL，并显示其稳定表达一种融合蛋白，该融合蛋白由（1）用于阳性T细胞选择的表面受体和（2）用于化疗后消除转染T细胞的“自杀”基因组成。本研究是首次报道表明SB转座子系统可介导人原代PBL中的稳定基因转移，这可能对基于T细胞的基因治疗有利。
