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DNA/Fusogenic Lipid Nanocarrier Assembly: Millisecond Structural Dynamics.

Borislav Angelov1, Angelina Angelova2, Sergey K Filippov1

  • 1†Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, CZ-16206 Prague, Czech Republic.

The Journal of Physical Chemistry Letters
|August 19, 2015
PubMed
Summary

This study reveals the millisecond-scale structural changes during DNA uptake by cationic lipid nanocarriers using time-resolved SAXS. These nanocarriers form lipoplexes with plasmid DNA, showing rapid structural transitions.

Keywords:
DNA-induced transitionlipoplexmillisecond kineticsself-assemblytime-resolved SAXS

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Area of Science:

  • Biophysics
  • Materials Science
  • Nanotechnology

Background:

  • Cationic lipid nanocarriers are crucial for gene delivery.
  • Understanding DNA complexation dynamics is key to optimizing delivery systems.
  • Cubosome nanoparticles offer unique nanochannel structures for cargo loading.

Purpose of the Study:

  • To investigate the ultrafast structural dynamics of DNA complexation with cationic lipid nanocarriers.
  • To characterize the structural transitions occurring during DNA uptake on a millisecond timescale.
  • To determine the rate of plasmid DNA (pDNA) complexation with lipid nanoparticles (NPs).

Main Methods:

  • Time-resolved small-angle X-ray scattering (SAXS) coupled with rapid-mixing stopped-flow technique.
  • Formation of cubosome nanoparticles from monoolein, DOMA, and DOPE-PEG2000.
  • Synchrotron X-ray beam for high-resolution structural analysis.
  • Dynamic roentgenogram recording at 4 ms time resolution.

Main Results:

  • Revealed ultrafast structural dynamics of cubosome particles complexing with neurotrophic plasmid DNA (pDNA).
  • Estimated the rate constant of pDNA/lipid NP complexation at millisecond resolution.
  • Observed a nanoparticle-nanoparticle structural transition and lipoplex formation upon pDNA upload.
  • Demonstrated tightly packed pDNA within the hydrated nanochannels of cubosome carriers.

Conclusions:

  • Cationic lipid nanocarriers undergo rapid structural changes during DNA uptake.
  • Cubosome structure facilitates efficient pDNA loading and lipoplex formation.
  • The study provides insights into the kinetics of gene carrier assembly.