Enhanced in vitro oligonucleotide and plasmid DNA transport by VP1 virus-like particles

Purpose. We developed a prokaryotic expression system to express the major capsid protein of Polyomavirus, VP1. Furthermore, we investigated the trans port of single stranded (ss) and double stranded (ds) DNA, mediated through VP1 as drug delivery system into mouse fibroblasts. Methods. To study DNA delivery we used two kinds of DNA, a ssDNA fragment ( 19mer) and dsDNA (plasmid pEGFPN1, 4.7 kb or a FITC-labelled dsDNA fragment , 1.8 kb). Results. The uptake of VP1 capsoids loaded with FITC-labelled oligodeoxynuc leotides (FODNs) was observed. VP1 pentamers loaded with condensates of den drimer/dsDNA fragments (FITC-labelled) resulted in significantly higher flu orescence signal in the cytoplasm of NIH 3T3 cells in comparison to control experiments without VP1. Additionally, VP1 capsoids loaded with plasmid pE GFPN1 without dendrimers resulted in an approximately 10 fold higher transf ection rate in comparison to blank DNA controls. Conclusions, Our results demonstrated the potential of VP1 capsoids as DNA delivery system. EGFP expression was significantly enhanced when plasmid DN A was delivered via VP1 capsoids, compared to control experiments with nake d DNA.