Quantifying the intracellular transport of viral and nonviral gene vectors in primary neurons

Exp Biol Med (Maywood). 2007 Mar;232(3):461-9.

Abstract

Real-time confocal particle tracking (CPT) was used to compare the transport and trafficking of polyethylenimine (PEI)/DNA nanocomplexes to that of efficient adenoviruses in live primary neurons. Surprisingly, the quantitative intracellular transport properties of PEI/DNA nonviral gene vectors are similar to that of adenoviral vectors. For example, the values of individual particle/virus transport rates and the distributions of particle/virus transport modes (i.e., the percentage undergoing active, diffusive, or subdiffusive transport) largely overlapped. In addition, both PEI/DNA vectors and adenoviruses rapidly accumulated near the cell nucleus in primary neurons despite our finding that PEI/DNA move slower in neurites than in the cell body, whereas adenoviruses move with equal rates in either location. The intracellular trafficking pathways of PEI/DNA and adenoviruses, however, were substantially different. The majority of PEI/DNA trafficked through the endolysosomal pathway so as to end up in late endosomes/lysosomes (LE/Lys), whereas adenoviruses efficiently escaped endosomes. This result suggests that the sequestration of nonviral gene vectors within acidic vesicles may be a critical barrier to gene delivery to primary neurons in the central nervous system (CNS).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adenoviridae / genetics
  • Adenoviridae / metabolism*
  • Animals
  • Biological Transport
  • Carboxylic Acids / chemistry
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • Endosomes / metabolism
  • Genetic Vectors / chemistry
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Lysosomes / metabolism
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Nanoparticles / chemistry
  • Neurites / metabolism
  • Neurons / cytology
  • Neurons / metabolism*
  • Polyethyleneimine / chemistry
  • Rats
  • Transfection

Substances

  • Carboxylic Acids
  • Oregon Green 488 carboxylic acid
  • Green Fluorescent Proteins
  • Polyethyleneimine