Murine leukemia virus vector integration favors promoter regions and regional hot spots in a human T-cell line
Murine leukemia virus vector integration favors promoter regions and regional hot spots in a human T-cell line
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DOI：
10.1016/j.bbrc.2006.05.007
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发表时间：
2006-07-07
影响因子：
3.1
通讯作者：
Takeshita, Toshikazu
中科院分区：
文献类型：
--
作者：
Tsukahara, Tomonori;Agawa, Hideyuki;Takeshita, Toshikazu
整合的基因组分析将是重要的，在评估安全性的人基因治疗与逆转录病毒载体。在这里，我们研究了人类T细胞中的MLV载体整合位点，因为它们适合基因转移研究，并且已在临床试验中用于治疗。我们在我们建立的感染的人T细胞克隆中定位了340个MLV载体整合位点。数据显示，MLV偏好在转录起始位点附近（+/-5 kb）、CpG岛附近（+/-1 kb）和RefSeq基因的第一个内含子内整合。我们还确定了在100 kb区域内包含三个或更多个整合的MLV整合热点。RT-PCR显示，与未感染的细胞相比，在转录起始位点或内含子附近含有MLV整合的T细胞克隆的mRNA水平失调。这些研究有助于确定MLV整合在T细胞中的概况以及与基于MLV的基因治疗相关的风险。(c)2006年爱思唯尔公司All rights reserved.
Genomic analysis of integration will be important in evaluating the safety of human gene therapy with retroviral vectors. Here, we investigated MLV vector integration sites in human T-cells, since they are amenable to gene transfer studies, and have been used therapeutically in clinical trials. We mapped 340 MLV vector integration sites in the infected human T-cell clones we established. The data showed that MLV preferred integration near the transcription start sites (+/- 5 kb), near CpG islands (+/- 1 kb), and within the first intron of RefSeq genes. We also identified MLV integration hot spots that contained three or more integrations within a 100 kb region. RT-PCR revealed that mRNA-levels of T-cell clones that contained MLV integrations near transcription start sites or introns were dysregulated compared to the uninfected cells. These studies help define the profile of MLV integration in T-cells and the risks associated with MLV-based gene therapy. (c) 2006 Elsevier Inc. All rights reserved.