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Excision of Sleeping Beauty transposons: parameters and applications to gene therapy
Geyi Liu; Elena L. Aronovich; Zongbin Cui; Chester B. Whitley; Perry B. Hackett; Hackett, PB, Discovery Genom Inc, 614 McKinley Pl NE, Minneapolis, MN 55413 USA
2004
Source PublicationJOURNAL OF GENE MEDICINE
Volume6Issue:5Pages:574-583
AbstractA 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.
KeywordFootprint Mouse Liver Tc1/mariner Terminal Repeats Transposition
DepartmentUniv 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
Subject AreaBiotechnology & Applied Microbiology ; Genetics & Heredity ; Medicine, Research & Experimental
Indexed BySCI
Language英语
WOS IDWOS:000221654100009
Citation statistics
Cited Times:59[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/9480
Collection期刊论文
Corresponding AuthorHackett, PB, Discovery Genom Inc, 614 McKinley Pl NE, Minneapolis, MN 55413 USA
Recommended Citation
GB/T 7714
Geyi Liu,Elena L. Aronovich,Zongbin Cui,et al. Excision of Sleeping Beauty transposons: parameters and applications to gene therapy[J]. JOURNAL OF GENE MEDICINE,2004,6(5):574-583.
APA Geyi Liu,Elena L. Aronovich,Zongbin Cui,Chester B. Whitley,Perry B. Hackett,&Hackett, PB, Discovery Genom Inc, 614 McKinley Pl NE, Minneapolis, MN 55413 USA.(2004).Excision of Sleeping Beauty transposons: parameters and applications to gene therapy.JOURNAL OF GENE MEDICINE,6(5),574-583.
MLA Geyi Liu,et al."Excision of Sleeping Beauty transposons: parameters and applications to gene therapy".JOURNAL OF GENE MEDICINE 6.5(2004):574-583.
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