RESUMO
In order to enhance the nuclear import of the transgene, we prepared plasmid DNA/importin-ß conjugates consisting of biotinylated poly(ethylenimine)s and recombinant streptavidin-fused importin-ß. Hemagglutinating virus of Japan-envelope vector containing the PEI polyplex/importin-ß conjugate showed high transfection efficiency not only in vitro but also in vivo. We showed that novel HVJ-E/importin-ß-conjugated PEI polyplex hybrid vector could overcome plasma and nuclear membrane barriers to achieve effective transfection.
Assuntos
DNA/administração & dosagem , Vetores Genéticos , Vírus Sendai/genética , beta Carioferinas/administração & dosagem , Animais , Biotinilação , Membrana Celular/metabolismo , Técnicas de Transferência de Genes , Masculino , Camundongos , Células NIH 3T3 , Membrana Nuclear/metabolismo , Plasmídeos , Polietilenoimina/química , Ratos , Ratos Wistar , Proteínas Recombinantes/química , Estreptavidina/química , Transfecção , Proteínas do Envelope Viral/genéticaRESUMO
In order to enhance the nuclear import of exogenous genes, novel plasmid DNA/importin-beta conjugates, which consist of a biotinylated plasmid DNA and a recombinant streptavidin-fused importin-beta, were prepared. The spacer length between plasmid DNA and biotin and the number of introduced biotin were adjusted. The microinjection of plasmid DNA/importin-beta conjugates into the cytoplasm of NIH3T3 cells resulted in the nuclear localization of conjugates and the higher expression efficiency, compared to intact plasmid DNA alone. These results indicate that plasmid DNA/importin-beta conjugates would be an important tool to enhance the nuclear localization of exogenous DNA in non-viral gene delivery system.
