Enhanced nuclear import and transfection efficiency of plasmid DNA using streptavidin-fused importin-beta.
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.
Nagasaki
Takeshi
T
Department of Applied and Bioapplied Chemistry, Graduate School of Engineering, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan. nagasaki@bioa.osaka-cu.ac.jp
Kawazu
Takeshi
T
Tachibana
Taro
T
Tamagaki
Seizo
S
Shinkai
Seiji
S
D021581:Active Transport, Cell Nucleus / Q000187:drug effects / Q000235:genetics; D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects / Q000235:genetics*; D004247:DNA / Q000008:administration & dosage / Q000235:genetics*; D020868:Gene Silencing; D051379:Mice; D008845:Microinjections; D041681:NIH 3T3 Cells; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000302:isolation & purification; D019809:Streptavidin / Q000008:administration & dosage / Q000235:genetics*; D014162:Transfection / Q000379:methods; D019076:Transgenes; D028961:beta Karyopherins / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics*
D011993:Recombinant Fusion Proteins; D028961:beta Karyopherins; D004247:DNA; D019809:Streptavidin