Journal article
Gene transfer into genomes of human cells by the sleeping beauty transposon system
Molecular therapy, Vol.8(1), pp.108-117
07/2003
DOI: 10.1016/S1525-0016(03)00099-6
PMID: 12842434
Abstract
The Sleeping Beauty (SB) transposon system, derived from teleost fish sequences, is extremely effective at delivering DNA to vertebrate genomes, including those of humans. We have examined several parameters of the SB system to improve it as a potential, nonviral vector for gene therapy. Our investigation centered on three features: the carrying capacity of the transposon for efficient integration into chromosomes of HeLa cells, the effects of overexpression of the SB transposase gene on transposition rates, and improvements in the activity of SB transposase to increase insertion rates of transgenes into cellular chromosomes. We found that SB transposons of about 6 kb retained 50% of the maximal efficiency of transposition, which is sufficient to deliver 70–80% of identified human cDNAs with appropriate transcriptional regulatory sequences. Overexpression inhibition studies revealed that there are optimal ratios of SB transposase to transposon for maximal rates of transposition, suggesting that conditions of delivery of the two-part transposon system are important for the best gene-transfer efficiencies. We further refined the SB transposase to incorporate several amino acid substitutions, the result of which led to an improved transposase called SB11. With SB11 we are able to achieve transposition rates that are about 100-fold above those achieved with plasmids that insert into chromosomes by random recombination. With the recently described improvements to the transposon itself, the SB system appears to be a potential gene-transfer tool for human gene therapy.
Details
- Title: Subtitle
- Gene transfer into genomes of human cells by the sleeping beauty transposon system
- Creators
- Aron M Geurts - Department of Genetics, Cell Biology and Development and The Arnold and Mabel Beckman Center for Transposon Research, University of Minnesota, Minneapolis, Minnesota 55455, USAYing Yang - Department of Genetics, Cell Biology and Development and The Arnold and Mabel Beckman Center for Transposon Research, University of Minnesota, Minneapolis, Minnesota 55455, USAKarl J Clark - Department of Genetics, Cell Biology and Development and The Arnold and Mabel Beckman Center for Transposon Research, University of Minnesota, Minneapolis, Minnesota 55455, USAGeyi Liu - Department of Genetics, Cell Biology and Development and The Arnold and Mabel Beckman Center for Transposon Research, University of Minnesota, Minneapolis, Minnesota 55455, USAZongbin Cui - Department of Genetics, Cell Biology and Development and The Arnold and Mabel Beckman Center for Transposon Research, University of Minnesota, Minneapolis, Minnesota 55455, USAAdam J Dupuy - Department of Genetics, Cell Biology and Development and The Arnold and Mabel Beckman Center for Transposon Research, University of Minnesota, Minneapolis, Minnesota 55455, USAJason B Bell - Department of Genetics, Cell Biology and Development and The Arnold and Mabel Beckman Center for Transposon Research, University of Minnesota, Minneapolis, Minnesota 55455, USADavid A Largaespada - Department of Genetics, Cell Biology and Development and The Arnold and Mabel Beckman Center for Transposon Research, University of Minnesota, Minneapolis, Minnesota 55455, USAPerry B Hackett - Department of Genetics, Cell Biology and Development and The Arnold and Mabel Beckman Center for Transposon Research, University of Minnesota, Minneapolis, Minnesota 55455, USA
- Resource Type
- Journal article
- Publication Details
- Molecular therapy, Vol.8(1), pp.108-117
- Publisher
- Elsevier Inc
- DOI
- 10.1016/S1525-0016(03)00099-6
- PMID
- 12842434
- ISSN
- 1525-0016
- eISSN
- 1525-0024
- Language
- English
- Date published
- 07/2003
- Academic Unit
- Anatomy and Cell Biology; Pathology
- Record Identifier
- 9984025692902771
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