Quantifying the intracellular transport of viral and nonviral gene vectors in primary neurons.
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).
Suk
Jung Soo
JS
Department of Chemical & Biomolecular Engineering, The Johns Hopkins University, Baltimore, MD 21218, USA.
Suh
Junghae
J
Lai
Samuel K
SK
Hanes
Justin
J
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D002264:Carboxylic Acids / Q000737:chemistry; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D011992:Endosomes / Q000378:metabolism; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D053758:Nanoparticles / Q000737:chemistry; D016501:Neurites / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D011094:Polyethyleneimine / Q000737:chemistry; D051381:Rats; D014162:Transfection
D002264:Carboxylic Acids; C436864:Oregon Green 488 carboxylic acid; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine