Example of Fatty Acid-Loaded Lipoplex in Enhancing in Vitro Gene Transfer Efficacies of Cationic Amphiphile

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 (lipid 1) whose in vitro gene transfer efficacies in CHO, COS-1, MCF-7, and HepG2 cells are remarkably enhanced wh...

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Bibliographic Details
Published in:Bioconjugate chemistry 2005-05, Vol.16 (3), p.676-684
Main Authors: Majeti, Bharat Kumar, Karmali, Priya Prakash, Madhavendra, Sunkara Sakunthala, Chaudhuri, Arabinda
Format: Article
Language:English
Subjects:
Animals
Cations - chemistry
Cell Line, Tumor
Cell Survival
Cercopithecus aethiops
Chemistry
Cricetinae
Electrophoretic Mobility Shift Assay
Fatty acids
Fatty Acids - chemistry
Genes
Humans
Ions
Liposomes - chemistry
Microscopy, Electron, Transmission
Molecular Structure
Transfection - instrumentation
Transfection - methods
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Summary: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 (lipid 1) whose in vitro gene transfer efficacies in CHO, COS-1, MCF-7, and HepG2 cells are remarkably enhanced when used in combination with 30 mole percent added myristic acid. Reporter gene expression assay using p-CMV-SPORT-β-gal reporter gene revealed poor gene transfer properties of the cationic liposomes of lipid 1 and cholesterol (colipid). However, the in vitro gene delivery efficacies of lipid 1 were found to be remarkably enhanced when the cationic liposomes of lipid 1 and cholesterol were prepared in the presence of 30 mole percent added myristic acid (with respect to lipid 1) as the third liposomal ingredient. The whole cell histochemical X-gal staining of representative CHO cells further confirmed the significantly enhanced gene transfer properties of the fatty acid-loaded cationic liposomes of lipid 1 and cholesterol. Electrophoretic gel patterns in the gel mobility shift assay supports the notion that better DNA release from fatty acid lipoplexes might play a role in their enhanced gene transfer properties. In addition, such myristic acid-loaded lipoplexes of lipid 1 were also found to be serum-compatible up to 30% added serum. Taken together, our present findings demonstrate that the transfection efficacies of fatty acid-loaded lipoplexes are worth evaluating particularly when traditional cationic liposomes prepared with either cholesterol or DOPE colipids fail to transfect cultured cells.
ISSN:1043-1802
1520-4812
DOI:10.1021/bc049687t