Optimization of methods to achieve mRNA-mediated transfection of tumor cells in vitro and in vivo employing cationic liposome vectors.
Direct in vivo transfection of tumor nodules in situ via liposome-DNA complexes has been employed as a strategy to accomplish antitumor immunization. To circumvent the potential safety hazards associated with systemic localization of delivered DNA, the utility of mRNA transcript-mediated gene delivery was explored. Capped, polyadenylated mRNA transcripts encoding the firefly luciferase and Escherichia coli lacZ reporter genes were derived by in vitro transcription. Transfection of the human breast cancer cell line MDA-MB-435 in vitro was accomplished employing cationic liposome-mRNA complexes. Evaluation of a panel of cationic liposome preparations demonstrated significant differences in the capacity of the various preparations to accomplish mRNA-mediated transfection. Quantitative evaluation of in vitro transfection demonstrated that target cells could be transfected at a high level of efficiency. The mRNA liposome-complexes were evaluated for in vivo transfection of tumor nodules in human xenografts in athymic nude mice. It could be demonstrated the liposome-mRNA complexes were comparable in efficacy to liposome-DNA complexes in accomplishing in situ tumor transfection. Thus, mRNA may be considered as an alternative to plasmid DNA as a gene transfer vector for genetic immunopotentiation applications.
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Gene Therapy Program, UAB Comprehensive Cancer Center, University of Alabama at Birmingham 35294, USA.
Benjamin
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Kim
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Conry
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RM
Curiel
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DT
D001943:Breast Neoplasms / Q000473:pathology*; D002411:Cation Exchange Resins; D002412:Cations; D004337:Drug Carriers; D005229:Fatty Acids, Monounsaturated; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005998:Glycine / Q000031:analogs & derivatives; D006801:Humans; D008055:Lipids; D008081:Liposomes; D008156:Luciferases / Q000096:biosynthesis; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D012333:RNA, Messenger / Q000008:administration & dosage / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012449:Safety; D013096:Spermine / Q000031:analogs & derivatives; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000096:biosynthesis
D002411:Cation Exchange Resins; D002412:Cations; D004337:Drug Carriers; D005229:Fatty Acids, Monounsaturated; D008055:Lipids; C086724:Lipofectamine; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C061134:dioctadecylamidoglycylspermine; D013096:Spermine; C020888:1,2-dielaidoylphosphatidylethanolamine; D008156:Luciferases; D001616:beta-Galactosidase; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane; D005998:Glycine