用于基因递送的肽TAT修饰的聚乙烯亚胺-β-环糊精
[Peptide TAT modified polyethylenimine-beta-cyclodextrin for gene delivery].
作者信息
Lai Li-Hua, Jiang Qi-Ying, Chen Dan, Hu Yi-Ping, Yu Hai, Wang Qing-Qing, Tang Gu-Ping
机构信息
Institute of Immunology, College of Medicine, Zhejiang University, Hangzhou 310058, China.
出版信息
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2009 Jan;38(1):15-23. doi: 10.3785/j.issn.1008-9292.2009.01.003.
OBJECTIVE
To develop a novel gene delivery vector TAT-PEI-beta-CyD.
METHODS
beta-cyclodextrin (beta-CyD) was linked by low molecular weight (PEI 600) via 1, 1-carbonyldiimidazole (CDI), and TAT peptide (RRRQRRKKRC) was coupled to PEI 600 by [N-succinimidy-3-(2-pyridyldithio) propionate, SPDP]. The copolymer was characterized by (1)H-NMR and FT-IR. Physiochemical characteristics of TAT-PEI-beta-CyD/DNA complexes were tested by agarose gel electrophoresis and particle size measurements. Cell viability and transfection efficiency were evaluated in A293 and B16 cells using PEI 25 kDa as a control.
RESULT
TAT peptide was successfully coupled to PEI-beta-CyD. The result of gel electrophoresis showed that the TAT-PEI-beta-CyD was able to condense DNA efficiently at N/P ratio of 4. The particle size of TAT-PEI-beta-CyD/DNA complexes was around 100 nm. The cytotoxicity of TAT-PEI-beta-CyD was lower than that of PEI 25 kDa. The transfection efficiency of TAT-PEI-beta-CyD was higher than that of PEI 25 kDa in A293 and B16 cells at N/P ratio of 30.
CONCLUSION
The novel vector TAT-PEI-beta-CyD has been developed successfully with low cytotoxicity and high transfection efficiency.
目的
研发一种新型基因递送载体TAT-PEI-β-环糊精。
方法
β-环糊精（β-CyD）通过1,1-羰基二咪唑（CDI）与低分子量聚乙烯亚胺（PEI 600）相连，TAT肽（RRRQRRKKRC）通过N-琥珀酰亚胺基-3-(2-吡啶二硫基)丙酸酯（SPDP）与PEI 600偶联。通过氢核磁共振（1H-NMR）和傅里叶变换红外光谱（FT-IR）对共聚物进行表征。采用琼脂糖凝胶电泳和粒度测量对TAT-PEI-β-CyD/DNA复合物的理化特性进行检测。以25 kDa聚乙烯亚胺作为对照，评估A293和B16细胞中的细胞活力和转染效率。
结果
TAT肽成功偶联至PEI-β-CyD。凝胶电泳结果表明，TAT-PEI-β-CyD在N/P比为4时能够有效凝聚DNA。TAT-PEI-β-CyD/DNA复合物的粒径约为100 nm。TAT-PEI-β-CyD的细胞毒性低于25 kDa聚乙烯亚胺。在N/P比为30时，TAT-PEI-β-CyD在A293和B16细胞中的转染效率高于25 kDa聚乙烯亚胺。
结论
成功研发出新型载体TAT-PEI-β-CyD，其具有低细胞毒性和高转染效率。
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