Abstract
Using live-cell confocal microscopy and particle tracking technology, the simultaneous transport of intracellular vesicles of the endo-lysosomal pathway and nonviral polyethylenimine (PEI)/DNA nanocomplexes was investigated. Due to potential problems associated with the use of acid-sensitive probes in combination with a gene vector that is hypothesized to buffer the pH of intracellular vesicles, the biological location of PEI/DNA gene vectors was revealed by probing their trafficking in cells expressing fluorescent versions of either early endosome antigen 1, a protein that localizes to early endosomes, or Niemann Pick C1, a protein that localizes to late endosomes and lysosomes. Studies directly show that PEI/DNA nanoparticles are actively transported within both early and late endosomes, and display similar overall transport rates in each. Additionally, gene vector transfer between endosomes is observed. Over time post-transfection, gene vectors accumulate in late endosomes/lysosomes; however, real-time escape of vectors from membrane-bound vesicles is not observed.
Original language | English (US) |
---|---|
Pages (from-to) | 691-697 |
Number of pages | 7 |
Journal | Microscopy Research and Technique |
Volume | 75 |
Issue number | 5 |
DOIs | |
State | Published - May 2012 |
Keywords
- Confocal microscopy
- Endosomes
- Lysosomes
- Particle tracking
- Polyethylenimine
ASJC Scopus subject areas
- Anatomy
- Histology
- Instrumentation
- Medical Laboratory Technology