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Efficient stable gene transfer into human cells by Sleeping Beauty transposon vectors

Item Type:Review
Title:Efficient stable gene transfer into human cells by Sleeping Beauty transposon vectors
Creators Name:Izsvak, Z. and Chuah, M.K. and VandenDriessche, T. and Ivics, Z.
Abstract:Transposable elements can be considered as natural, nonviral gene delivery vehicles capable of efficient genomic insertion. The plasmid-based transposon system of Sleeping Beauty (SB) combines the advantages of viruses and naked DNA molecules. In contrast to plasmid vectors, transposons integrate through a precise, recombinase-mediated mechanism into chromosomes, providing long-term expression of the gene of interest in cells. The advantages of transposons in comparison to viral systems include their simplicity and improved safety/toxicity profiles. In addition, the hyperactive SB100X is the first plasmid-based delivery system that overcomes the efficacy of non-viral delivery. The transposon delivery system consists of the transposase and the integration cassette, recognized by the transposase. The plasmid-based transposon delivery system can be combined with any non-viral delivery method. Here we provide two detailed protocols to apply SB-mediated, non-viral gene transfer in cultured cells. In our first example, we use a lipid-based delivery method in combination with the transposon-based integration system in an easy-to-transfect (HeLa) cell line. Second, we show how to achieve 40-50 % stable expression of a transgene in clinically relevant, hard-to-transfect cells (hematopoetic stem cells, HSCs) by nucleofection. The given protocols are adaptable to any vertebrate cells in culture.
Keywords:Sleeping Beauty, Transposon, Non-Viral, Gene Delivery, CD34+, HeLa, Gene Expression, Transfection, Nucleofection
Publisher:Elsevier / Academic Press
Page Range:287-297
Date:November 2009
Official Publication:https://doi.org/10.1016/j.ymeth.2009.07.001
PubMed:View item in PubMed

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