RNA interference in Vitro and in Vivo using a novel chitosan/siRNA nanoparticle system

Kenneth A Howard, Ulrik L Rahbek, Xiudong Liu, Christian Kroun Damgaard, Sys Zoffmann Glud, Morten Ø Andersen, Mads Bruun Hovgaard, Alexander Schmitz, Jens R Nyengaard, Flemming Besenbacher, Jørgen Kjems

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Abstract

This work introduces a novel chitosan-based siRNA nanoparticle delivery system for RNA interference in vitro and in vivo. The formation of interpolyelectrolyte complexes between siRNA duplexes (21-mers) and chitosan polymer into nanoparticles, ranging from 40 to 600 nm, was shown using atomic force microscopy and photon correlation spectroscopy. Rapid uptake (1 h) of Cy5-labeled nanoparticles into NIH 3T3 cells, followed by accumulation over a 24 h period, was visualized using fluorescence microscopy. Nanoparticle-mediated knockdown of endogenous enhanced green fluorescent protein (EGFP) was demonstrated in both H1299 human lung carcinoma cells and murine peritoneal macrophages (77.9% and 89.3% reduction in EGFP fluorescence, respectively). In addition, Western analysis showed approximately 90% reduced expression of BCR/ABL-1 leukemia fusion protein while BCR expression was unaffected in K562 (Ph(+)) cells after transfection using nanoparticles containing siRNA specific to the BCR/ABL-1 junction sequence. Effective in vivo RNA interference was achieved in bronchiole epithelial cells of transgenic EGFP mice after nasal administration of chitosan/siRNA formulations (37% and 43% reduction compared to mismatch and untreated control, respectively). These findings highlight the potential application of this novel chitosan-based system in RNA-mediated therapy of systemic and mucosal disease.
Original languageEnglish
JournalMolecular Therapy
Volume14
Issue4
Pages (from-to)476-484
Number of pages9
ISSN1525-0016
DOIs
Publication statusPublished - 2006

Keywords

  • Animals
  • Cells, Cultured
  • Chemistry, Physical
  • Chitosan
  • Genes, Reporter
  • Humans
  • Lung
  • Mice
  • Microscopy, Atomic Force
  • Nanostructures
  • Physicochemical Phenomena
  • RNA Interference
  • RNA, Small Interfering
  • Spectrum Analysis

Fingerprint

Dive into the research topics of 'RNA interference in Vitro and in Vivo using a novel chitosan/siRNA nanoparticle system'. Together they form a unique fingerprint.

Cite this