Cell-specific targeting by engineered M۱۳ bacteriophage expressing VEGFR۲ nanobody

Objective(s): Filamentous bacteriophage M۱۳ was genetically engineered to specifically target mammalian cells for gene delivery purpose. Materials and Methods: A vascular endothelial growth factor receptor ۲ (VEGFR۲)-specific nanobody was genetically fused to the capsid gene III of M۱۳ bacteriophage (pHEN۴/۳VGR۱۹). A mammalian expression construct containing Cop-green fluorescent protein (Cop-GFP), as a reporter gene, was amplified by PCR and then sub-cloned in the pHEN۴/۳VGR۱۹ phagemid. The resulting construct was transfected into ۲۹۳KDR cell. The recombinant phage was extracted and confirmed and then transduced into VEGFR۲ expressing cell (۲۹۳KDR). Results: Seventy-two hr after transfection, green fluorescence was detected in ۳۰% of the cells. About ۱% of the cells which transduced by recombinant phages were able to express GFP. Conclusion: It is hoped that the results from this study will help to find potential vectors to improve the efficiency of gene delivery. Taken together, we conclude that this newly-introduced vector can be used in cancer researches.

کلیدواژه ها:

Bacteriophage ، Nanobody ، Receptor-mediated gene transfer ، Targeted-gene delivery ، VEGFR۲