Mechanism Study of Gene Delivery and Expression in PK-15 Cells Using Magnetic Iron Oxide Nanoparticles as Gene Carriers

Abstract

The mechanism of gene delivery and expression is one of the most important concerns raised by the development of gene delivery methods. Limited investigation is performed on how magnetic nanoparticles combine with DNA and deliver gene into mammalian cells. In this context, polyethyleneimine (PEI) coated iron oxide magnetic nanoparticles (MNPs) were used as gene carriers for binding and condensing with plasmid DNA expressing enhanced green fluorescent protein (EGFP). The morphology and structure of MNP–DNA complexes were characterized by transmission electron microscopy (TEM) and atomic force microscopy (AFM). We evidenced that large amounts of DNA wrapped around the surface of MNPs and that the MNPs were physically entrapped by the DNA arranged both horizontally and vertically. EGFP gene was successfully expressed under mediation of an external magnetic field which is necessary to efficiently target EGFP gene to the cells. Fluorescence from EGFP was separately detected in the cell cytoplasm and cell nucleus.