Example of fatty acid-loaded lipoplex in enhancing in vitro gene transfer efficacies of cationic amphiphile

Majeti, Bharat Kumar ; Karmali, Priya Prakash ; Madhavendra, Sunkara Sakunthala ; Chaudhuri, Arabinda (2005) Example of fatty acid-loaded lipoplex in enhancing in vitro gene transfer efficacies of cationic amphiphile Bioconjugate Chemistry, 16 (3). pp. 676-684. ISSN 1043-1802

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Official URL: http://pubs.acs.org/doi/abs/10.1021/bc049687t

Related URL: http://dx.doi.org/10.1021/bc049687t

Abstract

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.

Item Type:Article
Source:Copyright of this article belongs to American Chemical Society.
ID Code:8028
Deposited On:25 Oct 2010 09:54
Last Modified:31 Jan 2011 08:30

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