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Simple derivation of transgene-free iPS cells by a dual recombinase approach

Item Type:Article
Title:Simple derivation of transgene-free iPS cells by a dual recombinase approach
Creators Name:Pertek, A. and Meier, F. and Irmler, M. and Beckers, J. and Skylaki, S. and Endele, M. and Wurst, W. and Prakash, N. and Kühn, R.
Abstract:Mammalian cells can be reprogrammed into induced pluripotent stem cells (iPSCs), a valuable tool for in vitro disease modeling and regenerative medicine. These applications demand for iPSCs devoid of reprogramming factor transgenes, but current procedures for the derivation of transgene-free iPSCs are inefficient and cumbersome. Here, we describe a new approach for the simple derivation of transgene-free iPSCs by the sequential use of two DNA recombinases, C31 Integrase and Cre, to control the genomic insertion and excision of a single, non-viral reprogramming vector. We show that such transgene-free iPSCs exhibit gene expression profiles and pluripotent developmental potential comparable to genuine, blastocyst-derived embryonic stem cells. As shown by a reporter iPSC line for the differentiation into midbrain dopaminergic neurons, the dual recombinase approach offers a simple and efficient way to derive transgene-free iPSCs for studying disease mechanisms and cell replacement therapies.
Keywords:iPS cells, Reprogramming, phiC31 Integrase, Cre Recombinase, Pluripotency, Animals, Mice
Source:Molecular Biotechnology
ISSN:1073-6085
Publisher:Springer / Humana Press (U.S.A.)
Volume:56
Number:8
Page Range:697-713
Date:August 2014
Official Publication:https://doi.org/10.1007/s12033-014-9748-y
PubMed:View item in PubMed

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