Generation of induced pluripotent stem cells from human foetal fibroblasts using the Sleeping Beauty transposon gene delivery system.
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.
Department of Anatomy & Embryology, Leiden University Medical Center, Leiden, The Netherlands; Netherlands Proteomics Institute, Utrecht, The Netherlands. Electronic address: r.p.davis@lumc.nl.
Richard P
RP
Davis
Csilla
C
Nemes
Eszter
E
Varga
Christian
C
Freund
Georgios
G
Kosmidis
Konstantinos
K
Gkatzis
Danielle
D
de Jong
Károly
K
Szuhai
András
A
Dinnyés
Christine L
CL
Mummery
D002454:Cell Differentiation; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D004251:DNA Transposable Elements / Q000235:genetics*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D018014:Gene Transfer Techniques; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism
D004251:DNA Transposable Elements
Differentiation; Human Induced pluripotent stem cell; Nuclear reprogramming; Sleeping Beauty transposon; Transfection methods