Example of Fatty Acid-Loaded Lipoplex in Enhancing in Vitro Gene Transfer Efficacies of Cationic Amphiphile
详细信息 查看全文
- 作者：Bharat Kumar Majeti ; Priya Prakash Karmali ; Sunkara Sakunthala Madhavendra ; and Arabinda Chaudhuri
- 刊名：Bioconjugate Chemistry
- 出版年：2005
- 出版时间：May 2005
- 年：2005
- 卷：16
- 期：3
- 页码：676 - 684
- 全文大小：299K
- 年卷期：v.16,no.3(May 2005)
- ISSN：1520-4812
文摘
Herein, we report on the design and synthesis of a novel nontoxic cationic amphiphile N,N-di-n-tetradecyl-N-[2-[N',N'-bis(2-hydroxyethyl)amino]ethyl]-N-(2-hydroxyethyl)ammonium chloride (lipid1) whose in vitro gene transfer efficacies in CHO, COS-1, MCF-7, and HepG2 cells are remarkablyenhanced when used in combination with 30 mole percent added myristic acid. Reporter geneexpression assay using p-CMV-SPORT-
-gal reporter gene revealed poor gene transfer properties ofthe cationic liposomes of lipid 1 and cholesterol (colipid). However, the in vitro gene delivery efficaciesof lipid 1 were found to be remarkably enhanced when the cationic liposomes of lipid 1 and cholesterolwere prepared in the presence of 30 mole percent added myristic acid (with respect to lipid 1) as thethird liposomal ingredient. The whole cell histochemical X-gal staining of representative CHO cellsfurther confirmed the significantly enhanced gene transfer properties of the fatty acid-loaded cationicliposomes of lipid 1 and cholesterol. Electrophoretic gel patterns in the gel mobility shift assay supportsthe notion that better DNA release from fatty acid lipoplexes might play a role in their enhancedgene transfer properties. In addition, such myristic acid-loaded lipoplexes of lipid 1 were also foundto be serum-compatible up to 30% added serum. Taken together, our present findings demonstratethat the transfection efficacies of fatty acid-loaded lipoplexes are worth evaluating particularly whentraditional cationic liposomes prepared with either cholesterol or DOPE colipids fail to transfectcultured cells.
-gal reporter gene revealed poor gene transfer properties ofthe cationic liposomes of lipid 1 and cholesterol (colipid). However, the in vitro gene delivery efficaciesof lipid 1 were found to be remarkably enhanced when the cationic liposomes of lipid 1 and cholesterolwere prepared in the presence of 30 mole percent added myristic acid (with respect to lipid 1) as thethird liposomal ingredient. The whole cell histochemical X-gal staining of representative CHO cellsfurther confirmed the significantly enhanced gene transfer properties of the fatty acid-loaded cationicliposomes of lipid 1 and cholesterol. Electrophoretic gel patterns in the gel mobility shift assay supportsthe notion that better DNA release from fatty acid lipoplexes might play a role in their enhancedgene transfer properties. In addition, such myristic acid-loaded lipoplexes of lipid 1 were also foundto be serum-compatible up to 30% added serum. Taken together, our present findings demonstratethat the transfection efficacies of fatty acid-loaded lipoplexes are worth evaluating particularly whentraditional cationic liposomes prepared with either cholesterol or DOPE colipids fail to transfectcultured cells.