Preparation of a transferrin-targeted M13-based gene nanocarrier and evaluation of its efficacy for gene delivery and expression in eukaryote cells

Abstract

Bacteriophages are appropriate gene carriers that might be targeted toward target cells using different strategies. Here we prepared a transferrin-targeted M13-based gene nanocarrier (Tf-targeted M13-GFP) and examined its gene delivery and expression efficacy in the AGS cell line. M13 phagemid particles bearing a GFP expression cassette (M13-GFP) were obtained from a recombinant lambda phage through an in vivo excision procedure. Chemical coupling of human holotransferrin molecules (Tf) to the surface of these phagemid particles resulted in Tf-targeted M13-GFP formation, which was then characterized by Phage-ELISA and Cell-ELISA experiments. Immunocytochemistry (ICC) and fluorescence-activated cell sorting (FACS) analysis were used for internalization assay and examination of gene delivery/expression efficacies in the human AGS cell line. The ELISA experiments revealed high-density attachment of Tf molecules to the surface of M13-GFP particles and ICC confirmed highly efficient internalization of the Tf-targeted M13-GFP particles into the AGS cells. Moreover, FACS analysis showed significant increase of GFP-positive cell counts in the samples treated with Tf-targeted M13-GFP (8.09%) in comparison with the samples treated with wild M13-GFP (1.2%). We conclude that this strategy might improve phage-mediated gene delivery and expression in eukaryote cells.