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Mol Biotechnol. 2014 Aug;56(8):697-713. doi: 10.1007/s12033-014-9748-y.

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

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  • 1Institute of Developmental Genetics, Helmholtz Zentrum München, 85764, Munich, Germany, anna.pertek@helmholtz-muenchen.de.

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.

PMID:
24677035
[PubMed - in process]
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