Messenger RNA electroporation is highly efficient in mouse embryonic stem cells

successful FLPe- and Cre-mediated recombination

P Ponsaerts, J P Brown, D Van den Plas, L Van den Eeden, D R Van Bockstaele, P G Jorens, V F I Van Tendeloo, J Merregaert, P B Singh, Z N Berneman

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Development of efficient short-term gene transfer technologies for embryonic stem (ES) cells is urgently needed for various existing and new ES cell-based research strategies. In this study, we present a highly efficient, nonviral non-DNA technology for genetic loading of mouse ES cells based on electroporation of defined mRNA. Here, we show that mouse ES cells can be efficiently loaded with mRNA encoding a green fluorescent reporter protein, resulting in a level of at least 90% of transgene expression without loss of cell viability and phenotype. To show that transgenes, introduced by mRNA electroporation, exert a specific cellular function in transfected cells, we electroporated stably transfected ES cell lines with mRNA encoding FLPe or Cre recombinase proteins in order to excise an FRT- or LoxP-flanked reporter gene. The results, as determined by the disappearance and/or appearance of a fluorescent reporter gene expression, show that FLPe and Cre recombinase proteins, introduced by mRNA electroporation, efficiently exert their function without influence on further culture of undifferentiated ES cell populations and their ability to differentiate towards a specific lineage.

Original languageEnglish
Pages (from-to)1606-10
Number of pages5
JournalGene Therapy
Volume11
Issue number21
DOIs
Publication statusPublished - Nov 2004

Fingerprint

Electroporation
Genetic Recombination
Embryonic Stem Cells
Messenger RNA
Transgenes
Reporter Genes
Technology Transfer
Stem Cell Research
Green Fluorescent Proteins
Cell Survival
Proteins
Mouse Embryonic Stem Cells
Technology
Phenotype
Gene Expression
Cell Line
Population
Genes

Keywords

  • Animals
  • Cells, Cultured
  • Electroporation
  • Flow Cytometry
  • Gene Expression
  • Green Fluorescent Proteins
  • Integrases
  • Luciferases, Firefly
  • Mice
  • RNA, Messenger
  • Recombination, Genetic
  • Stem Cells
  • Transgenes
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

Ponsaerts, P., Brown, J. P., Van den Plas, D., Van den Eeden, L., Van Bockstaele, D. R., Jorens, P. G., ... Berneman, Z. N. (2004). Messenger RNA electroporation is highly efficient in mouse embryonic stem cells: successful FLPe- and Cre-mediated recombination. Gene Therapy, 11(21), 1606-10. https://doi.org/10.1038/sj.gt.3302342

Messenger RNA electroporation is highly efficient in mouse embryonic stem cells : successful FLPe- and Cre-mediated recombination. / Ponsaerts, P; Brown, J P; Van den Plas, D; Van den Eeden, L; Van Bockstaele, D R; Jorens, P G; Van Tendeloo, V F I; Merregaert, J; Singh, P B; Berneman, Z N.

In: Gene Therapy, Vol. 11, No. 21, 11.2004, p. 1606-10.

Research output: Contribution to journalArticle

Ponsaerts, P, Brown, JP, Van den Plas, D, Van den Eeden, L, Van Bockstaele, DR, Jorens, PG, Van Tendeloo, VFI, Merregaert, J, Singh, PB & Berneman, ZN 2004, 'Messenger RNA electroporation is highly efficient in mouse embryonic stem cells: successful FLPe- and Cre-mediated recombination', Gene Therapy, vol. 11, no. 21, pp. 1606-10. https://doi.org/10.1038/sj.gt.3302342
Ponsaerts P, Brown JP, Van den Plas D, Van den Eeden L, Van Bockstaele DR, Jorens PG et al. Messenger RNA electroporation is highly efficient in mouse embryonic stem cells: successful FLPe- and Cre-mediated recombination. Gene Therapy. 2004 Nov;11(21):1606-10. https://doi.org/10.1038/sj.gt.3302342
Ponsaerts, P ; Brown, J P ; Van den Plas, D ; Van den Eeden, L ; Van Bockstaele, D R ; Jorens, P G ; Van Tendeloo, V F I ; Merregaert, J ; Singh, P B ; Berneman, Z N. / Messenger RNA electroporation is highly efficient in mouse embryonic stem cells : successful FLPe- and Cre-mediated recombination. In: Gene Therapy. 2004 ; Vol. 11, No. 21. pp. 1606-10.
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