Biotechnology Letters

, Volume 26, Issue 20, pp 1589–1592 | Cite as

Efficient gene transfer into murine embryonic stem cells by nucleofection

  • Peer Lorenz
  • Ulf Harnack
  • Rudolf Morgenstern

Abstract

Genetic manipulation of embryonic stem (ES) cells is performed by non-viral as well as viral transfection methods. We tested the recently developed nucleofection method delivering plasmid DNA directly into the nucleus for the introduction of a plasmid encoding enhanced green fluorescent protein (EGFP) into murine ES cells. Cell viability decreased from 77% before to 40% 24 h after nucleofection. Transfection effciencies in viable stem cells were between 85% and 96% with high levels of EGFP expression [mean fluorescence intensity (MFI): 630 ± 90] 24 h after nucleofection. After a two week culture in geneticin (G418) selection medium, nearly 50% of the stem cells were EGFP positive and continued transgene expression (MFIs: 120–240) for a two further weeks. We conclude that nucleofection is an efficient nonviral gene transfer method for the introduction of genes into murine ES cells.

gene transfer green fluorescent protein murine embryonic stem cells nucleofection 

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Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Peer Lorenz
    • 1
  • Ulf Harnack
    • 1
  • Rudolf Morgenstern
    • 1
  1. 1.Institute of Pharmacology and ToxicologyCharité Universitätsmedizin BerlinBerlinGermany

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