Cholesterol enhances cationic liposome-mediated DNA transfection of human respiratory epithelial cells

Author:

Bennett Michael J.1,Nantz Michael H.1,Balasubramaniam Rajiv P.1,Gruenert Dieter C.2,Malone Robert W.3

Affiliation:

1. Gene Therapy Program, Department of Chemistry, University of California, Davis, California 95616

2. Gene Therapy Core Center, Cystic Fibrosis Research Center and Cardiovascular Research Institute, Department of Laboratory Medicine, University of California, San Francisco, California 94143

3. Medical Pathology, University of California, Davis, California 95616

Abstract

Cationic liposome transfection reagents are useful for transferring polynucleotides into cells, and have been proposed for human pulmonary gene therapy. The effect of adding cholesterol to cationic lipid preparations has been tested by first formulating the cationic lipid N-[1-(2,3-dioleoyloxy)propyl-N-[1-(2-hydroxy)ethyl]-N,N-dimethyl ammonium iodide (DORI) with varying amounts of dioleoylphos-phatidylethanolamine (DOPE) and cholesterol. Cholesterol was found to enhance lipid-mediated transfection in both the respiratory epithelial cells and mouse fibroblasts. These findings will facilitate nucleic acid transfection of many cell types including differentiated epithelial cell monolayers, and therefore may be useful for examining gene regulation in various cell types and for developing pulmonary gene therapy.

Publisher

Portland Press Ltd.

Subject

Cell Biology,Molecular Biology,Biochemistry,Biophysics

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