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学科主题: Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental
题名: Excision of Sleeping Beauty transposons: parameters and applications to gene therapy
作者: Geyi Liu ; Elena L. Aronovich ; Zongbin Cui ; Chester B. Whitley ; Perry B. Hackett
通讯作者: Hackett, PB, Discovery Genom Inc, 614 McKinley Pl NE, Minneapolis, MN 55413 USA
关键词: footprint ; mouse liver ; Tc1/mariner ; terminal repeats ; transposition
刊名: JOURNAL OF GENE MEDICINE
发表日期: 2004
卷: 6, 期:5, 页:574-583
收录类别: SCI
部门归属: Univ Minnesota, Dept Genet Cell Biol & Dev, Minneapolis, MN 55455 USA; Univ Minnesota, Inst Human Genet, Minneapolis, MN 55455 USA; Univ Minnesota, Arnold & Mabel Beckman Ctr Transposon Res, Minneapolis, MN 55455 USA; Univ Minnesota, Dept Pediat, Minneapolis, MN USA; Chinese Acad Sci, Inst Hydrobiol, Wuhan, Peoples R China; Discovery Genom Inc, Minneapolis, MN 55413 USA
摘要: A major problem in gene therapy is the determination of the rates at which gene transfer has occurred. Our work has focused on applications of the Sleeping Beauty (SB) transposon system as a non-viral vector for gene therapy. Excision of a transposon from a donor molecule and its integration into a cellular chromosome are catalyzed by SB transposase. In this study, we used a plasmid-based excision assay to study the excision step of transposition. We used the excision assay to evaluate the importance of various sequences that border the sites of excision inside and outside the transposon in order to determine the most active sequences for transposition from a donor plasmid. These findings together with our previous results in transposase binding to the terminal repeats suggest that the sequences in the transposon-junction of SB are involved in steps subsequent to DNA binding but before excision, and that they may have a role in transposase-transposon interaction. We found that SB transposons leave characteristically different footprints at excision sites in different cell types, suggesting that alternative repair machineries operate in concert with transposition. Most importantly, we found that the rates of excision correlate with the rates of transposition. We used this finding to assess transposition in livers of mice that were injected with the SB transposon and transposase. The excision assay appears to be a relatively quick and easy method to optimize protocols for delivery of genes in SB transposons to mammalian chromosomes in living animals. Copyright (C) 2004 John Wiley Sons, Ltd.
语种: 英语
WOS记录号: WOS:000221654100009
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/9480
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

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.Excision of Sleeping Beauty transposons: parameters and applications to gene therapy,JOURNAL OF GENE MEDICINE,2004,6(5):574-583
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