Dioleoylmelittin as a novel serum-insensitive reagent for efficient transfection of mammalian cells
| Autor: | Andreas Supersaxo, Jean-Yves Legendre, Bohrmann B, Arnold Trzeciak, Ulrich Deuschle, Kitas E |
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| Rok vydání: | 1997 |
| Předmět: |
Biomedical Engineering
Pharmaceutical Science Bioengineering Peptide Gene delivery Transfection Hemolysis Melittin Fatty Acids Monounsaturated chemistry.chemical_compound Dogs Amphiphile Animals Pharmacology chemistry.chemical_classification Gel electrophoresis Membranes Phosphatidylethanolamines Organic Chemistry DNA Melitten Quaternary Ammonium Compounds Microscopy Electron chemistry Biochemistry Solubility Covalent bond COS Cells Liposomes lipids (amino acids peptides and proteins) Indicators and Reagents Biotechnology Plasmids |
| Zdroj: | Bioconjugate chemistry. 8(1) |
| ISSN: | 1043-1802 |
| Popis: | Amphipathic peptides can be useful effectors to enhance gene delivery. However, peptide/DNA complexes usually require additional effectors, such as fusogenic lipids, to mediate efficient transfection. Due to weak and/or multiple interactions between the various components of the system, the transfecting complexes are often heterogeneous and unstable in biological fluids. Accordingly, a hybrid molecule resulting from the covalent coupling of an amphipathic, membrane-disturbing peptide to a lipid moiety might create a stable and efficient peptide-based gene transfer system. The present work describes such a novel hybrid molecule, dioleoylmelittin, resulting from the conjugation of dioleoylphosphatidylethanolamine-N-[3-(2-pyridyldithio)propionate] with [Cys1]melittin. Dioleoylmelittin had a lower hemolytic and membrane-disturbing activity than melittin. Size and zeta potential measurements, DNA gel electrophoresis, and electron microscopy showed that dioleoylmelittin, unlike melittin, was able to complex plasmid DNA to form spherical particles with a net positive charge and a diameter between 50 and 250 nm. These particles, prepared at an optimal 10/1 dioleoylmelittin/DNA ratio (w/w), mediated efficient transient transfection of reporter genes in cultured mammalian cells including primary cells. The luciferase activity induced by the dioleoylmelittin/DNA complex was 5-500-fold higher than that induced by a cationic lipid/DNA complex, depending on the cationic lipid and the cell-line. Surprisingly, the presence of 10-50% fetal calf serum during dioleoylmelittin-mediated transfection enhanced 1.5-3-fold gene expression. Dioleoylmelittin represents a new class of efficient peptide-based transfection reagents, especially suited for serum-sensitive cells. |
| Databáze: | OpenAIRE |
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