Generation of induced pluripotent stem cells from human foetal fibroblasts using the Sleeping Beauty transposon gene delivery system

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• 作者：Richard P. Davis ; Csilla Nemes ; Eszter Varga ; Christian Freund ; Georgios Kosmidis ; Konstantinos Gkatzis ; Danielle de Jong ; K¨¢roly Szuhai ; Andr¨¢s Dinny¨¦s ; Christine L. Mummery
• 关键词：Human Induced pluripotent stem cell ; Sleeping Beauty transposon ; Differentiation ; Transfection methods ; Nuclear reprogramming
• 刊名：Differentiation
• 出版年：2013
• 出版时间：July-September, 2013
• 年：2013
• 卷：86
• 期：1-2
• 页码：30-37
• 全文大小：6644 K

文摘

Transposon gene delivery systems offer an alternative, non-viral-based approach to generate induced pluripotent stem cells (iPSCs). Here we used the Sleeping Beauty (SB) transposon to generate four human iPSC lines from foetal fibroblasts. In contrast to other gene delivery systems, the SB transposon does not exhibit an integration bias towards particular genetic elements, thereby reducing the risk of insertional mutagenesis. Furthermore, unlike the alternative transposon piggyBac, SB has no SB-like elements within the human genome, minimising the possibility of mobilising endogenous transposon elements. All iPSC lines exhibited the expected characteristics of pluripotent human cells, including the ability to differentiate to derivatives of all three germ layers in vitro. Re-expression of the SB transposase in the iPSCs after reprogramming resulted in the mobilisation of some of the transposons. These results indicate that the SB transposon system is a useful addition to methods for generating human iPSCs, both for basic and applied biomedical research, and in the context of future therapeutic application.