title: Real-time gene delivery vector tracking in the endo-lysosomal pathway of live cells.
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.
authors.affiliation: Department of Bioengineering, Rice University, 6100 Main St. MS-142, Houston, Texas 77005, USA. jsuh@rice.edu
authors.forename: Junghae
authors.initials: J
authors.lastname: Suh
authors.forename: Yoojin
authors.initials: Y
authors.lastname: An
authors.forename: Benjamin C
authors.initials: BC
authors.lastname: Tang
authors.forename: Christopher
authors.initials: C
authors.lastname: Dempsey
authors.forename: Feiran
authors.initials: F
authors.lastname: Huang
authors.forename: Justin
authors.initials: J
authors.lastname: Hanes
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D011992:Endosomes / Q000737:chemistry* / Q000378:metabolism; D018014:Gene Transfer Techniques; D008247:Lysosomes / Q000737:chemistry* / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D053758:Nanoparticles / Q000032:analysis*; D013997:Time Factors
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