Journal article
Optimization of cationic lipid-mediated gene transfer to airway epithelia
The American journal of physiology, Vol.269(1 Pt 1), pp.L45-L51
07/1995
DOI: 10.1152/ajplung.1995.269.1.L45
PMID: 7631813
Abstract
The use of cationic lipids for gene transfer to airway epithelia has shown promise in in vitro and in vivo studies. However, previous studies have used a wide variety of different lipid preparations and different formulations. Few studies have been designed to optimize the variables involved in transfection, and none have been focused on airway epithelia. Therefore we examined variables that affect cationic lipid-mediated transfection of HeLa cells and of airway epithelial cells grown on permeable filter supports at the air-liquid interface. To quantitate expression of cDNA, we assayed expression of luciferase. We found that the ratio of DNA to lipid was an important variable that determined transfection efficiency. In both HeLa and airway epithelial cells, the optimum charge ratio of cationic lipid to anionic DNA was approximately 1.25, consistent with the notion that a positively charged complex facilitates interaction with the negatively charged cell membrane. After testing a series of readily available cationic lipids, we found that 1,2-dimyristyloxypropyl-N,N-dimethyl-hydroxyethyl ammonium bromide (DMRIE)/dioleoyl phosphatidylethandamine (DOPE) appeared to show good efficacy. The concentration of DNA and cationic lipid also played an important role: in HeLa cells the optimum concentration of cationic lipid was approximately 5 microM and in airway epithelial cells was approximately 15 microM.
Details
- Title: Subtitle
- Optimization of cationic lipid-mediated gene transfer to airway epithelia
- Creators
- A J Fasbender - Howard Hughes Medical Institute, Department of Internal Medicine, University of Iowa College of Medicine, Iowa City 52242, USAJ ZabnerM J Welsh
- Resource Type
- Journal article
- Publication Details
- The American journal of physiology, Vol.269(1 Pt 1), pp.L45-L51
- DOI
- 10.1152/ajplung.1995.269.1.L45
- PMID
- 7631813
- NLM abbreviation
- Am J Physiol
- ISSN
- 0002-9513
- eISSN
- 2163-5773
- Publisher
- American Physiological Society; United States
- Grant note
- HL-51670 / NHLBI NIH HHS HL-50257 / NHLBI NIH HHS DK-25295 / NIDDK NIH HHS
- Language
- English
- Date published
- 07/1995
- Academic Unit
- Neurology; Molecular Physiology and Biophysics; Pulmonary, Critical Care, and Occupational Medicine; Fraternal Order of Eagles Diabetes Research Center; Neurosurgery; Internal Medicine
- Record Identifier
- 9984020861302771
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