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Conventional plasmid vectors (pDNA) possess unwanted prokaryotic DNA that when delivered to the mammalian cells can reduce transgene expression and integrate into the human genome, potentiating oncogenesis. Bacterial sequence-free, linear covalently closed (LCC) DNA minivectors have shown improved safety and efficacy both in vitro and in vivo. We previously constructed and characterized an in vivo LCC DNA minivector (ministring) system that can convert pDNA into DNA ministrings. Here we sought to optimize this system and apply ministrings to encode a gene to generate HIV virus-like particles (VLPs) in targeted cells. The DNA-VLP expresses gagV3(BCE) gene in target cells to simultaneously yield GagV3(BCE) VLP assembly and presentation of immunodominant HIV antigens, eliciting production of neutralizing antibodies and cytotoxic T-cell responses against HIV.

Optimized production of a safe and efficient gene therapeutic vaccine versus HIV via a linear covalently closed DNA minivector