Enhanced in vitro oligonucleotide and plasmid DNA transport by VP1 virus-like particles.
OBJECTIVE
We developed a prokaryotic expression system to express the major capsid protein of Polyomavirus, VP1. Furthermore, we investigated the transport 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 oligodeoxynucleotides (FODNs) was observed. VP1 pentamers loaded with condensates of dendrimer/dsDNA fragments (FITC-labelled) resulted in significantly higher fluorescence signal in the cytoplasm of NIH 3T3 cells in comparison to control experiments without VP1. Additionally, VP1 capsoids loaded with plasmid pEGFPN1 without dendrimers resulted in an approximately 10 fold higher transfection 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 DNA was delivered via VP1 capsoids, compared to control experiments with naked DNA.
Henke
S
S
Institute for Pharmaceutical Technology, Biocenter-Johann Wolfgang Goethe-University, Frankfurt am Main, Germany.
Rohmann
A
A
Bertling
W M
WM
Dingermann
T
T
Zimmer
A
A
D016475:3T3 Cells; D000818:Animals; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D016503:Drug Delivery Systems / Q000379:methods; D051379:Mice; D016376:Oligonucleotides, Antisense / Q000235:genetics* / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D011120:Polyomavirus / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D004268:DNA-Binding Proteins; D016376:Oligonucleotides, Antisense; D010940:Plant Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D004247:DNA