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DNA-membrane complex formation during electroporation is DNA size-dependent.

Shaurya Sachdev1, Sara Feijoo Moreira1, Yasmine Keehnen1

  • 1Department of Chemical Engineering, Delft University of Technology, van der Maasweg 9, Delft 2629 HZ, The Netherlands.

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Summary

DNA molecule size impacts cell membrane interaction during electroporation. Larger DNA molecules (≥25 base pairs) form aggregates, while smaller ones (15 base pairs) do not, affecting intracellular transport.

Keywords:
DNA aggregationDNA sizeDNA-membrane complex formationElectropermeabilizationElectroporationGene electrotransfer

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Area of Science:

  • Biophysics
  • Molecular Biology
  • Cell Biology

Background:

  • Electroporation is a method for introducing molecules into cells.
  • The size of DNA molecules is a critical factor influencing their behavior during cellular processes.
  • Understanding DNA-membrane interactions is key to optimizing gene delivery and other biotechnological applications.

Purpose of the Study:

  • To investigate the effect of DNA molecule size on DNA-membrane interactions during electroporation.
  • To determine the threshold size for DNA aggregate formation at the cell membrane.
  • To analyze how DNA size influences translocation into the cytoplasm.

Main Methods:

  • Cells were subjected to electroporation with various DNA molecule sizes (15, 25, 50, 100, and 1000 base pairs).
  • Microscopy and biophysical techniques were employed to observe DNA-membrane complex and aggregate formation.
  • Quantification of DNA translocation into the cytoplasm was performed for each size.

Main Results:

  • DNA molecules of 25 base pairs or larger formed observable DNA-membrane complexes or aggregates.
  • No aggregation was observed for 15 base pair DNA molecules.
  • Direct cytoplasmic access was achieved for all tested DNA sizes, but the amount of translocated DNA decreased as molecule size increased.
  • The size-dependent aggregation phenomenon aligns with Onsager's condensation theory for rod-like molecules.

Conclusions:

  • DNA size critically dictates its interaction with the cell membrane during electroporation, influencing aggregate formation.
  • A minimum size of 25 base pairs is required for DNA to form aggregates at the cell membrane under the tested conditions.
  • While electroporation facilitates cytoplasmic entry for all DNA sizes, larger molecules exhibit reduced translocation efficiency, a finding consistent with theoretical predictions.