Eom Khee Dong, Park Sun Mi, Tran Huu Dung, Kim Myong Soo, Yu Ri Na, Yoo Hoon
Nanormics Inc., Sungbuk-gu, Seoul, 136-130, South Korea.
Pharm Res. 2007 Aug;24(8):1581-9. doi: 10.1007/s11095-006-9231-y. Epub 2007 Mar 21.
To evaluate the potential use of dendritic alpha,epsilon-poly(L-lysine)s (DPL) for the efficient cellular delivery of antisense oligonucleotides.
A series of dendritic alpha,epsilon-polylysines of various generations were prepared. Their physical properties and the ability to form complex with oligonucleotide were investigated by polyacrylamide gel electrophoresis, capillary zone electrophoresis (CZE), agarose gel electrophoresis, fluorescence titration and atomic force microscopy (AFM). The efficiency to deliver oligonucleotide to HeLa cells, stably transfected with plasmid pLuc/705, was evaluated by using antisense splicing correction assay and confocal microscopy.
DPLs formed the complexes with antisense oligonucleotide with modest cytotoxicity. The charge ratio of oligonucleotide to DPL and the size (generation) of DPLs were all critical variables for the antisense effect. Compared to low generation DPLs, high generation DPLs were more effective in delivering oligonucleotide into cells.
High generation DPL-oligonucleotide complexes were moderately effective for delivery antisense oligonucleotide. The complex formation provides a promise for in vivo therapeutic application of DPLs or their derivatives in the delivery of gene or oligonucleotide.
评估树枝状α,ε-聚（L-赖氨酸）（DPL）在反义寡核苷酸高效细胞递送中的潜在用途。
制备了一系列不同代数的树枝状α,ε-聚赖氨酸。通过聚丙烯酰胺凝胶电泳、毛细管区带电泳（CZE）、琼脂糖凝胶电泳、荧光滴定和原子力显微镜（AFM）研究了它们的物理性质以及与寡核苷酸形成复合物的能力。使用反义剪接校正试验和共聚焦显微镜评估了将寡核苷酸递送至稳定转染质粒pLuc/705的HeLa细胞中的效率。
DPL与反义寡核苷酸形成复合物，具有适度的细胞毒性。寡核苷酸与DPL的电荷比以及DPL的大小（代数）都是影响反义效果的关键变量。与低代数DPL相比，高代数DPL在将寡核苷酸递送至细胞中更有效。
高代数DPL-寡核苷酸复合物在递送反义寡核苷酸方面具有中等效果。复合物的形成有望使DPL或其衍生物在基因或寡核苷酸递送的体内治疗应用中发挥作用。
