Delivery of short interfering ribonucleic acid-complexed magnetic nanoparticles in an oscillating field occurs via caveolae-mediated endocytosis

PLoS One. 2012;7(12):e51350. doi: 10.1371/journal.pone.0051350. Epub 2012 Dec 7.

Abstract

Gene delivery technologies to introduce foreign genes into highly differentiated mammalian cells have improved significantly over the last few decades. Relatively new techniques such as magnetic nanoparticle-based gene transfection technology are showing great promise in terms of its high transfection efficiency and wide-ranging research applications. We have developed a novel gene delivery technique, which uses magnetic nanoparticles moving under the influence of an oscillating magnetic array. Herein we successfully introduced short interfering RNA (siRNA) against green fluorescent protein (GFP) or actin into stably-transfected GFP-HeLa cells or wild-type HeLa and rat aortic smooth muscle cells, respectively. This gene silencing technique occurred in a dose- and cell density- dependent manner, as reflected using fluorescence intensity and adhesion assays. Furthermore, using endocytosis inhibitors, we established that these magnetic nanoparticle-nucleic acid complexes, moving across the cell surface under the influence of an oscillating magnet array, enters into the cells via the caveolae-mediated endocytic pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caveolae / metabolism*
  • Endocytosis / physiology*
  • Flow Cytometry
  • Gene Transfer Techniques*
  • Green Fluorescent Proteins
  • HeLa Cells
  • Humans
  • Magnetic Fields*
  • Magnetite Nanoparticles / administration & dosage*
  • RNA, Small Interfering / administration & dosage*
  • Rats
  • Tetrazolium Salts
  • Thiazoles

Substances

  • Magnetite Nanoparticles
  • RNA, Small Interfering
  • Tetrazolium Salts
  • Thiazoles
  • Green Fluorescent Proteins
  • thiazolyl blue

Grants and funding

This research is wholly funded by nanoTherics Limited. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.