Non-degradative intracellular trafficking of highly compacted polymeric DNA nanoparticles

Anthony J. Kim, Nicholas J. Boylan, Jung Soo Suk, Samuel K. Lai, Justin Hanes

Research output: Contribution to journalArticle

Abstract

Highly compacted DNA nanoparticles (DNPs) composed of polyethylene glycol linked to a 30-mer of poly-l-lysine via a single cysteine residue (CK 30PEG) have previously been shown to provide efficient gene delivery to the brain, eyes and lungs. In this study, we used a combination of flow cytometry, high-resolution live-cell confocal microscopy, and multiple particle tracking (MPT) to investigate the intracellular trafficking of highly compacted CK 30PEG DNPs made using two different molecular weights of PEG, CK 30PEG 10k and CK 30PEG 5k. We found that PEG MW did not have a major effect on particle morphology nor nanoparticle intracellular transport. CK 30PEG 10k and CK 30PEG 5k DNPs both entered human bronchial epithelial (BEAS-2B) cells via a caveolae-mediated pathway, bypassing degradative endolysosomal trafficking. Both nanoparticle formulations were found to rapidly accumulate in the perinuclear region of cells within 2 h, 37 ± 19% and 47 ± 8% for CK 30PEG 10k and CK 30PEG 5k, respectively. CK 30PEG 10k and CK 30PEG 5k DNPs moved within live cells at average velocities of 0.09 ± 0.04 μm/s and 0.11 ± 0.04 μm/s, respectively, in good agreement with reported values for caveolae. These findings show that highly compacted DNPs employ highly regulated trafficking mechanisms similar to biological pathogens to target specific intracellular compartments.

Original languageEnglish (US)
Pages (from-to)102-107
Number of pages6
JournalJournal of Controlled Release
Volume158
Issue number1
DOIs
StatePublished - Feb 28 2012

Keywords

  • Cystic fibrosis
  • Cystic fibrosis
  • Gene therapy
  • Gene therapy
  • Intracellular trafficking
  • Intracellular trafficking
  • Nonviral
  • Nonviral
  • Particle tracking
  • Particle tracking

ASJC Scopus subject areas

  • Pharmaceutical Science

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