Open Access Green as soon as Postprint is submitted to ZB.
Simple derivation of transgene-free iPS cells by a dual recombinase approach.
Mol. Biotechnol. 56, 697-713 (2014)
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.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Cre Recombinase ; Ips Cells ; Phic31 Integrase ; Pluripotency ; Reprogramming; Pluripotent Stem-cells; Site-specific Integration; Somatic-cells; Mammalian-cells; Dopaminergic-neurons; Human Fibroblasts; Phic31 Integrase; Phage Integrase; Gene-expression; Generation