用于基因递送的肽CY11偶联聚乙烯亚胺-β-环糊精
[Peptide CY11 conjugated polyethylenimine-beta-cyclodextrin for gene delivery].
作者信息
Hu Yi-Ping, Jiang Qi-Ying, Chen Dan, Zhao Dan-Jun, Lu Xiao, Yu Hai, Wang Qing-Qing, Zhou Jun, Hu Xiu-Rong, Tang Gu-Ping
机构信息
Institute of Chemical Biology and Pharmaceutical Chemistry, Zhejiang University, Hangzhou 310028, China.
出版信息
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2009 Jan;38(1):24-30. doi: 10.3785/j.issn.1008-9292.2009.01.004.
OBJECTIVE
To develop a novel non-viral gene delivery vector CY11-PEI-beta-CyD and to test its gene transfection efficiency.
METHODS
CY11 (CGMQLPLATWY) was conjugated to polyethylenimine-beta-cyclodextrin to form CY11-PEI-beta-CyD with a cross-linker [N-succinimidy-3-(2-pyridyldithio) propionate, SPDP]. (1)H-NMR and TGA were used to confirm the structure of vector. The DNA condensing ability of CY11-PEI-beta-CyD was investigated by gel retardation assay. Cytotoxicity of CY11-PEI-beta-CyD was determined by MTT assay and transfection efficiency was investigated in COS-7, Hela and B16 cells.
RESULT
CY11 was conjugated onto PEI-beta-CyD successfully, confirmed by(1)H NMR and TGA. The novel vector effectively condensed DNA at N/P ratio of 4îIt showed low cytotoxicity up to the concentration was 160 Mgr;g/ml. The transfection efficiency was 17-fold higher than that of PEI 25 kDa at N/P ratio of 20.
CONCLUSION
The novel vector CY11 -PEI-beta-CyD with low cytotoxic and high transfection efficiency may be used as a potential carrier for gene delivery.
目的
开发一种新型非病毒基因递送载体CY11-PEI-β-环糊精，并测试其基因转染效率。
方法
将CY11（CGMQLPLATWY）与聚乙烯亚胺-β-环糊精通过交联剂[N-琥珀酰亚胺基-3-（2-吡啶二硫基）丙酸酯，SPDP]偶联形成CY11-PEI-β-环糊精。利用核磁共振氢谱（1H-NMR）和热重分析（TGA）确认载体结构。通过凝胶阻滞试验研究CY11-PEI-β-环糊精的DNA凝聚能力。采用MTT法测定CY11-PEI-β-环糊精的细胞毒性，并在COS-7、Hela和B16细胞中研究其转染效率。
结果
通过1H NMR和TGA证实CY11成功偶联到PEI-β-环糊精上。该新型载体在N/P比为4时能有效凝聚DNA；在浓度高达160 μg/ml时显示出低细胞毒性。在N/P比为20时，其转染效率比25 kDa的聚乙烯亚胺（PEI）高17倍。
结论
新型载体CY11-PEI-β-环糊精具有低细胞毒性和高转染效率，可作为一种潜在的基因递送载体。
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