Transgene-free production of pluripotent stem cells using piggyBac transposons

Knut Woltjen, Riikka Hämäläinen, Mark Kibschull, Maria Mileikovsky, Andras Nagy

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

49 Citations (Scopus)

Abstract

Reprogramming of somatic cells to induced pluripotent stem cells (iPSCs) allows the derivation of -personalized stem cells. Transposon transgenesis is a novel and viable alternative to viral transduction methods for the delivery of reprogramming factors (Oct4, Sox2, Klf4, c-Myc) to somatic cells. Since transposons can be introduced as naked DNA using common plasmid transfection protocols, they provide a safer alternative to viral methods. piggyBac transposons are host-factor independent and integrate stably into the target genome, yet benefit from the unique characteristic of seamless removal mediated by transient expression of piggyBac transposase. Thus, piggyBac transposition provides an effective means to generate human, transgene-free iPSCs. The protocol describes the production of iPSCs from human embryonic fibroblasts, delivering reprogramming factors via plasmid transfection and piggyBac transposition.

Original languageEnglish
Title of host publicationHuman Pluripotent Stem Cells
Subtitle of host publicationMethods and Protocols
Pages87-103
Number of pages17
Volume767
DOIs
Publication statusPublished - 2011
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume767
ISSN (Print)1064-3745

Keywords

  • dox
  • doxycycline regulated
  • factor-free
  • induced pluripotent stem cells
  • iPS cells
  • iPSC
  • nonviral
  • piggyBac
  • reprogramming
  • transfection
  • transgene removal
  • transgene-free
  • transposition
  • transposon
  • virus-free

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