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90plus 納米粒度儀應用案例-17
閱讀：637 發布時間：2014-3-1790plus 納米粒度儀應用案例
文獻名：Direct Gene Transfer to the Respiratory Tract of Mice with Pure Plasmid and Lipid-Formulated DNA
作者：MICHAEL J. McCLUSKIE,1,2 YONGLIANG CHU,5 JIU-LIN XIA, 5 JOEL JESSEE, 5 GULILAT GEBYEHU, 5 and HEATHER L. DAVIS1,2,3,4
1Loeb Research Institute, Ottawa, Canada.
2department of Cellular and Molecular Medicine,
3Department of Microbiology and Immunology, Faculty of Medicine,
4School of Rehabilitation Sciences, Faculty of Health Sciences, University of Ottawa, Ottawa, Canada.
5Life Technologies, Inc., Rockville, MD 20850.
摘要：Direct gene transfer into the respiratory system could be carried out for either therapeutic or immunization purposes. Here we demonstrate that cells in the lung can take up and express plasmid DNA encoding a luciferase reporter gene whether it is administered in naked form or formulated with cationic liposomes. Depending on the lipid used, the transfection efficiency with liposome-formulated DNA may be higher, the same as, or less than that with pure plasmid DNA. Tetramethyltetraalkylspermine analogs with alkyl groups of 16 or 18 carbons and DMRIE/cholesterol formulations proved particularly effective. Similar results for reporter gene expression in the lung were obtained whether the DNA (naked or lipid formulated) was administered by indirect, noninvasive intranasal delivery (inhaled or instilled) or by invasive, direct intratracheal delivery (injected or via a cannula). Reporter gene expression peaks around 4 days, then falls off dramatically by 9 days. The dose-response is linear, at least up to 100 μg plasmid DNA, suggesting better transfection efficiencies might be realized if there was not a volume limitation. For a given dose of DNA, the best results are obtained when the DNA is mixed with the minimum amount of lipid that can complex it compley. These results are discussed in the context of direct gene transfer for either gene therapy or delivery of a mucosal DNA vaccine.