Yoo Hyuk Sang, Jeong Seo Young
Department of Biomaterials Engineering, Kangwon National University, Chuncheon, Republic of Korea.
Eur J Pharm Biopharm. 2007 Apr;66(1):28-33. doi: 10.1016/j.ejpb.2006.09.013. Epub 2006 Oct 10.
A nuclear localization signal was non-covalently attached to DNA for the purpose of enhancing transfection efficiencies of non-viral gene carriers. Psoralen, a nucleic acid-intercalating agent, was chemically attached to a signal peptide. The conjugate spontaneously intercalated into DNA and then poly(ethyleneimine) [PEI] was added to prepare a DNA/PEI complex containing the signal peptide moieties. The existence of the conjugate did not alter the complexation process between DNA and PEI, which was confirmed by dynamic light scattering. The conjugate was slowly released from the DNA/PEI complex for 24h, while a burst release was examined when the conjugated was added to DNA without PEI. The complex containing a signal peptide moiety increased transfection efficiencies on COS-1 cells, compared to a mutant signal peptide or a control. Cytotoxicity of the conjugate slowly increased as the amount of the conjugate increased, however, the cytotoxic effect of the conjugate was not significant at the effective concentration of the conjugate for transfections. Therefore, the psoralen-nuclear localization signal is expected to be a potent transfection enhancing agent without a covalent modification of transgenes.
为提高非病毒基因载体的转染效率，将核定位信号非共价连接到DNA上。补骨脂素是一种核酸嵌入剂，化学连接到信号肽上。该共轭物自发嵌入DNA，然后加入聚乙烯亚胺（PEI）以制备含有信号肽部分的DNA/PEI复合物。动态光散射证实，共轭物的存在并未改变DNA与PEI之间的复合过程。共轭物从DNA/PEI复合物中缓慢释放24小时，而当共轭物加入不含PEI的DNA中时，则观察到突发释放。与突变信号肽或对照相比，含有信号肽部分的复合物提高了对COS-1细胞的转染效率。随着共轭物数量的增加，共轭物的细胞毒性缓慢增加，然而，在转染的共轭物有效浓度下，共轭物的细胞毒性作用并不显著。因此，补骨脂素-核定位信号有望成为一种有效的转染增强剂，而无需对转基因进行共价修饰。
