Real-time gene delivery vector tracking in the endo-lysosomal pathway of live cells

Junghae Suh, Yoojin An, Benjamin C. Tang, Christopher Dempsey, Feiran Huang, Justin Hanes

Research output: Contribution to journalArticle

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 languageEnglish (US)
Pages (from-to)691-697
Number of pages7
JournalMicroscopy Research and Technique
Volume75
Issue number5
DOIs
StatePublished - May 1 2012

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Keywords

  • Confocal microscopy
  • Endosomes
  • Lysosomes
  • Particle tracking
  • Polyethylenimine

ASJC Scopus subject areas

  • Anatomy
  • Histology
  • Instrumentation
  • Medical Laboratory Technology

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