A novel miniaturized filamentous phagemid as a gene delivery vehicle to target mammalian cells

The filamentous phage M13 is a single-stranded DNA phage with several attractive characteristics for gene delivery, including a capsid amenable to the display of foreign peptides and a simple well-characterized genome that is easy to genetically modify. Previously, we constructed a DNA minivector based on M13 (a miniphagemid), which minimized the inflammatory bacterial and phage DNA content in the vector. In general, DNA minivectors devoid of their prokaryotic components have shown improved gene transfer and safety. We examined the miniphagemid's capacity for <i>in vitro</i> transgene delivery to target cells through phage display of epidermal growth factor to target its cognate receptor. The absence of the prokaryotic backbone and smaller vector size conferred by the miniphagemids were associated with improved transgene expression for purified single-stranded phagemid DNA and phagemid virion particles. We further engineered this system to enhance packaging of DNA minivectors via deletion of the packaging signal within the helper plasmid used to produce miniphagemids and observed improved phage-mediated gene expression in mammalian cells. Overall, we present a set of novel transgene delivery vectors that combine cell-targeting ligand display and vector minimization. This platform showcases the flexibility of M13 as a gene delivery tool with immense therapeutic potential.