A multifunctional envelope-type nano device for novel gene delivery of siRNA plasmids.
A multifunctional envelope-type nano device (MEND) for use in the delivery of siRNA expression plasmids is described. The plasmid DNA encoding anti-luciferase short interfering RNA (siRNA) was condensed by poly-L-lysine (PLL) and packaged into the MEND. The silencing effect of the MEND(PLL) showed a 96% inhibition of luciferase activities in a co-transfection study. The silencing effect was maintained at more than 60%, even under the 100-fold diluted conditions. In the luciferase transformed cells, however, the MEND(PLL) showed no significant silencing effect (10%), indicating heterogeneity in transfection by the MEND(PLL). To solve this problem, the DNA condensing agents were optimized by comparing PLL, stearyl octaarginine (STR-R8) and protamine (Prot). No difference in silencing effect (95-97%) was found among these MENDs in a co-transfection study. However, the MEND(Prot) showed a 70% silencing effect in the transformed cells. These results suggest that the MEND(Prot) has less heterogeneity in transfection, while the MEND(PLL) and the MEND(STR-R8) have large heterogeneities. These results demonstrate that MEND(Prot) is a promising gene delivery system for siRNA expression plasmids with less heterogeneity associated with the transfection.
Moriguchi
Rumiko
R
Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo 060-0812, Japan.
Kogure
Kentaro
K
Akita
Hidetaka
H
Futaki
Shiroh
S
Miyagishi
Makoto
M
Taira
Kazunari
K
Harashima
Hideyoshi
H
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004339:Drug Compounding; D004587:Electrophoresis, Agar Gel; D005079:Excipients; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D008156:Luciferases / Q000235:genetics; D049329:Nanostructures; D009842:Oligopeptides / Q000737:chemistry; D010957:Plasmids / Q000235:genetics; D011479:Protamines / Q000737:chemistry; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics*; D013228:Stearates / Q000737:chemistry; D014162:Transfection
D005079:Excipients; D009842:Oligopeptides; D011479:Protamines; D034741:RNA, Small Interfering; D013228:Stearates; C487882:stearyl-octaarginine; D008156:Luciferases