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Nanosecond electric pulse effects on gene expression.

Louise Chopinet1, Tina Batista-Napotnik, Audrey Montigny

  • 1CNRS, IPBS - UMR 5089, 205 route de Narbonne, BP 64182, 31077, Toulouse Cedex 4, France.

The Journal of Membrane Biology
|July 9, 2013
PubMed
Summary
This summary is machine-generated.

Nanosecond electric pulses (nsEPs) were investigated to enhance gene electrotransfer efficiency. However, nsEPs did not improve plasmid delivery to the nucleus or green fluorescent protein expression in CHO cells.

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Molecular Biology

Background:

  • Gene electrotransfer using micro/millisecond electric pulses is a standard gene delivery method.
  • Efficient gene transfer requires plasmid DNA to reach the cell nucleus.
  • Nanosecond electric pulses (nsEPs) may enhance nuclear uptake by affecting internal cell membranes.

Purpose of the Study:

  • To evaluate the efficacy of nsEPs in augmenting gene electrotransfer.
  • To determine if nsEPs improve plasmid DNA delivery to the nucleus.
  • To assess the impact of nsEPs on gene expression and constitutive protein levels.

Main Methods:

  • Chinese hamster ovary (CHO) cells were subjected to classical gene electrotransfer followed by nsEPs.
  • Various time gaps (0.5-24 h) and nsEP parameters (18-60 kV/cm) were tested.
  • Green fluorescent protein (GFP) expression was analyzed using fluorescence microscopy and flow cytometry.

Main Results:

  • nsEPs did not increase the percentage of electrotransfected CHO cells.
  • The amount of GFP expressed in CHO cells was not enhanced by nsEP application.
  • nsEPs showed no effect on constitutive GFP fluorescence intensity in HCT-116 cells.

Conclusions:

  • nsEPs do not significantly contribute to gene electrotransfer in CHO cells.
  • nsEPs do not impact constitutive GFP expression in HCT-116 cells.
  • Further research may be needed to explore alternative applications of nsEPs in cell biology.