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

Authors : Mohammad KHALAJKONDORI, Mahboobeh KAVOOSI, Mohammad RAHMATIYAMCHI, Mehdi KADIVAR

Pages : 561-570

View : 15 | Download : 10

Publication Date : 2016-12-01

Article Type : Research Paper

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 insert ignore into journalissuearticles values(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 insert ignore into journalissuearticles values(M13-GFP); were obtained from a recombinant lambda phage through an in vivo excision procedure. Chemical coupling of human holotransferrin molecules insert ignore into journalissuearticles values(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 insert ignore into journalissuearticles values(ICC); and fluorescence-activated cell sorting insert ignore into journalissuearticles values(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 insert ignore into journalissuearticles values(8.09%); in comparison with the samples treated with wild M13-GFP insert ignore into journalissuearticles values(1.2%);. We conclude that this strategy might improve phage-mediated gene delivery and expression in eukaryote cells.
Keywords : Chemical coupling, phage mediated gene transfer, gene delivery, transferrin targeted delivery