Gateway compatible transposon vector to genetically modify human embryonic kidney and adipose-derived stromal cells

Item Type:	Article
Title:	Gateway compatible transposon vector to genetically modify human embryonic kidney and adipose-derived stromal cells
Creators Name:	Petrakis, S. and Rasko, T. and Mates, L. and Ivics, Z. and Izsvak, Z. and Kouzi-Koliakou, K. and Koliakos, G.
Abstract:	The Gateway Technology cloning system and the transposon technology represent state-of-the-art laboratory techniques. Combination of these molecular tools may allow rapid cloning of target genes into expression vectors. Here, we describe a novel Gateway Technology compatible transposon plasmid that combines the advantages of the Gateway recombination cloning with the Sleeping Beauty (SB) transposon-mediated transgene integrations. In our system the transposition is catalyzed by the novel hyperactive SB100x transposase providing the high efficient and precise transgene integrations into the host genome. A Gateway compatible transposon plasmid was generated in which the potential target gene may be fused with a Yellow Fluorescent Protein (YFP) tag at the N'- terminal. The vector utilizes the CAGGS promoter to control the fusion protein expression. The transposon expression vector encoding fusion YFP-IFNB1 protein together with the hyperactive SB100x transposase was used to generate stable cell lines in human embryonic kidney (HEK293) and rat adipose-derived stromal cells (ASC). HEK293 and ASCs stably expressed and secreted human interferon-beta protein up to 4 weeks after transfection. The generated Gateway compatible transposon plasmid may be utilized for numerous experimental approaches, such as gene therapy or high-throughput screening methods in primary cells, representing a valuable molecular tool for laboratory applications.
Keywords:	Methods, Gene Expression, Transposon, Gateway, Animals, Rats
Source:	Biotechnology Journal
ISSN:	1860-6768
Publisher:	Wiley
Volume:	7
Number:	7
Page Range:	891-897
Date:	July 2012
Official Publication:	https://doi.org/10.1002/biot.201100471
PubMed:	View item in PubMed

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