In vitro gene transfection using dendritic poly(L-lysine).
Monodispersed dendritic poly(L-lysine)s (DPKs) of several generations were synthesized, and their characteristics as a gene transfection reagent were then investigated. The agarose gel shift and ethidium bromide titration assay proved that the DPKs of the third generation and higher could form a complex with a plasmid DNA, and the degree of compaction of the DNA was increased by the increasing number of the generation. The DPKs of the fifth and sixth generation, which have 64 and 128 amine groups on the surface of the molecule, respectively, showed efficient gene transfection ability into several cultivated cell lines without significant cytotoxity. In addition, the transfection efficiency of the DPK of the sixth generation was not seriously reduced even if serum was added at 50% of the final concentration into the transfection medium. Because we can strictly synthesize various DPK derivatives, which have several types of branch units, terminal cationic groups, and so on, they are expected to be a good object of study regarding the basic information on the detailed mechanism of gene transfection into cells. We also expect to be able to easily construct DPK-based functional gene carriers, e.g., DPKs modified by ligands such as a sugar chain, which can enable advanced gene delivery in vivo.
Ohsaki
Mio
M
Department of Applied Chemistry, Faculty of Engineering, Nagasaki University, 852-8521, Japan.
Okuda
Tatsuya
T
Wada
Akihiro
A
Hirayama
Toshiya
T
Niidome
Takuro
T
Aoyagi
Haruhiko
H
D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D019556:COS Cells; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured / Q000378:metabolism*; D002850:Chromatography, Gel; D006224:Cricetinae; D004247:DNA / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids; D011107:Polylysine / Q000235:genetics*; D014162:Transfection / Q000379:methods*
D011107:Polylysine; D004247:DNA