Reprogramming of human fibroblasts to pluripotent stem cells using mRNA of four transcription factors

Eduard Yakubov, Gidi Rechavi, Shmuel Rozenblatt, David Givol

Research output: Contribution to journalArticle

162 Scopus citations

Abstract

Reprogramming of differentiated cells into induced pluripotent cells (iPS) was accomplished in 2006 by expressing four, or less, embryonic stem cell (ESC)-specific transcription factors. Due to the possible danger of DNA damage and the potential tumorigenicity associated with such DNA damage, attempts were made to minimize DNA integration by the vectors involved in this process without complete success. Here we present a method of using RNA transfection as a tool for reprogramming human fibroblasts to iPS. We used RNA synthesized in vitro from cDNA of the same reprogramming four transcription factors. After transfection of the RNA, we show intracellular expression and nuclear localization of the respective proteins in at least 70% of the cells. We used five consecutive transfections to support continuous protein expression resulting in the formation of iPS colonies that express alkaline phosphatase and several ESC markers and that can be expanded. This method completely avoids DNA integration and may be developed to replace the use of DNA vectors in the formation of iPS.

Original languageEnglish (US)
Pages (from-to)189-193
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume394
Issue number1
DOIs
StatePublished - Mar 26 2010

Keywords

  • DNA integration
  • iPS
  • Malignancy
  • Reprogramming
  • RNA transfection
  • Transcription factors

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

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