Gene delivery in three-dimensional cell cultures by superparamagnetic nanoparticles

Haiyuan Zhang, Moo Yeal Lee, Michael G. Hogg, Jonathan S. Dordick, Susan T. Sharfstein

Research output: Contribution to journalArticlepeer-review

81 Scopus citations

Abstract

Three-dimensional (3D) cellular assays closely mimic the in vivo milieu, providing a rapid, inexpensive system for screening drug candidates for toxicity or efficacy in the early stages of drug discovery. However, 3D culture systems may suffer from mass transfer limitations, particularly in delivery of large polypeptide or nucleic acid compounds. Nucleic acids (e.g., genes, silencing RNA) are of particular interest both as potential therapeutics and due to a desire to modulate the gene-expression patterns of cells exposed to small-molecule pharmacological agents. In the present study, polyethylenimine (PEI)-coated superparamagnetic nanoparticles (SPMNs) were designed to deliver interfering RNA and green fluorescent protein (GFP) plasmids through a collagen-gel matrix into 3D cell cultures driven by an external magnetic field. The highest transfection efficiency achieved was 64% for siRNA and 77% for GFP plasmids. Delivery of an shRNA plasmid against GFP by PEI-coated SPMNs silenced the GFP expression with 82% efficiency. We further demonstrated that this delivery approach could be used for screening interfering RNA constructs for therapeutic or toxic effects for cells grown in 3D cultures. Four known toxic shRNA plasmids were delivered by PEI-coated SPMNs into 3D cell cultures, and significant toxicities (41-51% cell death) were obtained.

Original languageEnglish
Pages (from-to)4733-4743
Number of pages11
JournalACS Nano
Volume4
Issue number8
DOIs
StatePublished - 24 Aug 2010
Externally publishedYes

Keywords

  • Cytotoxicity
  • Gene expression
  • Gene transfer
  • Magnetism
  • Silencing RNA

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