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

Authors

  • A. Pertek
  • F. Meier
  • M. Irmler
  • J. Beckers
  • S. Skylaki
  • M. Endele
  • W. Wurst
  • N. Prakash
  • R. Kühn

Journal

  • Molecular Biotechnology

Citation

  • Mol Biotechnol 56 (8): 697-713

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.


DOI

doi:10.1007/s12033-014-9748-y