Structural advantage of dendritic poly(L-lysine) for gene delivery into cells.
This study aimed to investigate the relationships between structures of gene carrier molecules and their activities for gene delivery into cells. We compared 2 types of poly(L-lysine) as carriers, that is, dendritic poly(L-lysine) (KG6) and linear poly(L-lysine) (PLL). KG6 formed a neutral DNA complex, and its DNA compaction level was weaker than that of PLL. The amount of DNA binding and uptake into cells mediated by PLL was 4-fold higher than that with KG6. However, KG6-mediated gene expression was 100-fold higher than that by PLL. Since pK(a) values of terminal amines of KG6 were lowered even though small amounts of DNA were internalized into cells, sufficient DNA amounts for effective gene expression escaped to the cytosol due to the proton sponge effect in the endosome. In addition, weakly compacted DNA with KG6 was advantageous in accessing RNA polymerase in the cell nucleus. On the other hand, PLL did not show the proton sponge effect in the endosome and resulted in strong compaction of DNA. Even though large DNA amounts were internalized into cells, most of the DNA would not take part in gene expression systems in the nucleus. Amount of induced cytokine production after intravenous injection of DNA complexes with KG6 and PLL was low, and was similar to the case when DNA was injected alone. Therefore, no significant difference in effects on cytokine production was observed between KG6 and PLL.
Yamagata
Masato
M
Department of Applied Chemistry, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.
Kawano
Takahito
T
Shiba
Kouhei
K
Mori
Takeshi
T
Katayama
Yoshiki
Y
Niidome
Takuro
T
D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D016207:Cytokines / Q000494:pharmacology; D004247:DNA / Q000737:chemistry / Q000187:drug effects; D050091:Dendrimers / Q000737:chemistry*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D018014:Gene Transfer Techniques; D008156:Luciferases / Q000187:drug effects / Q000235:genetics; D015394:Molecular Structure; D011107:Polylysine / Q000737:chemistry*; D013329:Structure-Activity Relationship
D016207:Cytokines; D050091:Dendrimers; D011107:Polylysine; D004247:DNA; D008156:Luciferases