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Nature Reviews Molecular Cell Biology volume 10, page 238 (2009)Cite this article

Reprogramming of somatic cells to a pluripotent state, thereby creating induced pluripotent stem (iPS) cells, requires the transient expression of one or more transcription factors. The efficiency of adenoviral and plasmid transfection to deliver reprogramming transcription factor genes is very low, and neither method has been applied successfully to human cells. Three new reports describe the non-viral transfection of mouse and human cells using a transposable element, called piggyBac (PB), which carries one or more transcription factor genes and inserts itself into cellular DNA.

The PB system requires inverted terminal repeats that flank the transgene cassette, and transient expression of the transposase enzyme to catalyse insertion and excision events. Efforts led by Woltjen et al. and by Kaji et al. resulted in the efficient reprogramming of mouse and human embryonic fibroblasts, which expressed characteristic pluripotency markers, indicating a reprogrammed state. This was confirmed by in vitro differentiation assays and chimaera development. The reprogramming cassette was efficiently excised from mouse cells by transient expression of transposase or Cre recombinase; yet, the pluripotent state was stably maintained. Guo et al. reprogrammed partly specialized mouse cells, called epistem cells, into iPS cells using a single reprogramming factor, KLF4. Following Cre-mediated excision of the Klf4 transgene, the iPS cells maintained themselves using the endogenous Klf4 gene, which had been switched on during reprogramming. So, reprogramming factors can be removed without a trace from iPS cells once exogenous expression becomes dispensable.

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ORIGINAL RESEARCH PAPERS

  1. Woltjen, K. et al. piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells. Nature 1 Mar 2009 (doi:10.1038/nature07863)

    Article  CAS  Google Scholar 

  2. Kaji, K. et al. Virus-free induction of pluripotency and subsequent excision of reprogramming factors. Nature 1 Mar 2009 (doi:10.1038/nature07864)

    Article  CAS  Google Scholar 

  3. Guo, G. et al. Klf4 reverts developmentally programmed restriction of ground state pluripotency. Development 136, 1063–1069 (2009)

    Article  CAS  Google Scholar 

  4. Eilken, H. M. et al. Continuous single-cell imaging of blood generation from haemogenic endothelium. Nature 457, 896–900 (2009)

    Article  CAS  Google Scholar 

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Technology watch. Nat Rev Mol Cell Biol 10, 238 (2009). https://doi.org/10.1038/nrm2664

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