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首页> 外文OA文献 >Gene Transfer Efficiency and Genome-Wide Integration Profiling of Sleeping Beauty, Tol2, and PiggyBac Transposons in Human Primary T Cells
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Gene Transfer Efficiency and Genome-Wide Integration Profiling of Sleeping Beauty, Tol2, and PiggyBac Transposons in Human Primary T Cells

机译：人类原代T细胞中睡美人，Tol2和PiggyBac转座子的基因转移效率和全基因组整合分析

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In this study, we compared the genomic integration efficiencies and transposition site preferences of Sleeping Beauty (SB or SB11), Tol2, and piggyBac (PB) transposon systems in primary T cells derived from peripheral blood lymphocytes (PBL) and umbilical cord blood (UCB). We found that PB demonstrated the highest efficiency of stable gene transfer in PBL-derived T cells, whereas SB11 and Tol2 mediated intermediate and lowest efficiencies, respectively. Southern hybridization analysis demonstrated that PB generated the highest number of integrants when compared to SB and Tol2 in both PBL and UCB T cells. Tol2 and PB appeared more likely to promote clonal expansion than SB, which may be in part due to the dysregulated expression of cancer-related genes near the insertion sites. Genome-wide integration analysis demonstrated that SB, Tol2, and PB integrations occurred in all the chromosomes without preference. Additionally, Tol2 and PB integration sites were mainly localized near transcriptional start sites (TSSs), CpG islands and DNaseI hypersensitive sites, whereas SB integrations were randomly distributed. These results suggest that SB may be a preferential choice of the delivery vector in T cells due to its random integration site preference and relatively high efficiency, and support continuing development of SB-mediated T-cell phase I trials.
机译：在这项研究中，我们比较了来自外周血淋巴细胞（PBL）和脐带血（UCB）的原代T细胞中Sleeping Beauty（SB或SB11），Tol2和piggyBac（PB）转座子系统的基因组整合效率和转座位点偏好）。我们发现PB证明了在PBL衍生的T细胞中稳定基因转移的最高效率，而SB11和Tol2分别介导了中等效率和最低效率。 Southern杂交分析表明，与PBL和UCB T细胞中的SB和Tol2相比，PB产生的整合子数量最多。 Tol2和PB似乎比SB更有可能促进克隆扩增，这可能部分是由于插入位点附近的癌症相关基因表达失调所致。全基因组范围的整合分析表明，SB，Tol2和PB整合在所有染色体上均无优先发生。此外，Tol2和PB整合位点主要位于转录起始位点（TSS），CpG岛和DNaseI超敏位点附近，而SB整合则是随机分布的。这些结果表明，由于其随机整合位点偏好和相对较高的效率，SB可能是T细胞中递送载体的优先选择，并支持SB介导的T细胞I期试验的持续发展。

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