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

Anna Pertek, Florian Meier, Martin Irmler, Johannes Beckers, Stavroula Skylaki, Max Endele, Wolfgang Wurst, Nilima Prakash, Ralf Kühn

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


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.

Original languageEnglish
Pages (from-to)697-713
Number of pages17
JournalApplied Biochemistry and Biotechnology - Part B Molecular Biotechnology
Issue number8
StatePublished - Aug 2014


  • Cre recombinase
  • Pluripotency
  • Reprogramming
  • iPS cells
  • phiC31 integrase


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