Novel dextran–spermine conjugates as transfecting agents: comparing water-soluble and micellar polymers
| العنوان: | Novel dextran–spermine conjugates as transfecting agents: comparing water-soluble and micellar polymers |
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| المؤلفون: | Aviva Joseph, A. Makovitzki, Tony Azzam, Abraham J. Domb, Dan Gazit, Hagit Eliyahu, A Zlotkin, Yechezkel Barenholz |
| المصدر: | Gene Therapy. 12:494-503 |
| بيانات النشر: | Springer Science and Business Media LLC, 2004. |
| سنة النشر: | 2004 |
| مصطلحات موضوعية: | Cytoplasm, Polymers, viruses, Genetic enhancement, Cell Culture Techniques, Gene Expression, Golgi Apparatus, Spermine, Culture Media, Serum-Free, Blood substitute, Fatty Acids, Monounsaturated, Mice, chemistry.chemical_compound, Polyethyleneimine, Transgenes, Luciferases, Micelles, Zebrafish, chemistry.chemical_classification, Mice, Inbred C3H, Microscopy, Confocal, Dextrans, Polymer, Flow Cytometry, Dextran, Biochemistry, Molecular Medicine, Female, lipids (amino acids, peptides, and proteins), Intracellular, Plasmids, animal structures, macromolecular substances, Gene delivery, Biology, Transfection, Cell Line, Injections, Cations, Genetics, Animals, Humans, Molecular Biology, Cell Nucleus, technology, industry, and agriculture, Genetic Therapy, Culture Media, Quaternary Ammonium Compounds, chemistry, NIH 3T3 Cells, Conjugate |
| الوصف: | Recently, a novel cationic polymer, dextran-spermine (D-SPM) was developed for gene delivery. An efficient transfection was obtained using this polycation for a variety of genes and cell lines in serum-free or serum-poor medium. However, transfection using the water-soluble D-SPM-based polyplexes decreased with increasing serum concentration in cell culture in a concentration-dependent manner, reaching 95% inhibition at 50% serum in the cell growth medium. In order to overcome this obstacle, oleyl derivatives of D-SPM (which form micelles in aqueous phase) were synthesized at 1, 10, and 20 mol% of oleyl moiety to polymer epsilon-NH2 to form N-oleyl-D-SPM (ODS). Polyplexes based on ODS transfected well in medium containing 50% serum. Comparison with polyplexes based on well-established polymers (branched and linear polyethyleneimine) and with DOTAP/Cholesterol lipoplexes showed that regarding beta-galactosidase transgene expression level and cytotoxicity in tissue culture, the D-SPM and ODS compare well with the above polyplexes and lipoplexes. Intracellular trafficking using FITC-labeled ODS and Rhodamine-labeled pGeneGrip plasmid cloned with hBMP2 monitored by confocal microscopy revealed that during the transfection process the fluorescent-labeled polymer concentrates in the Golgi apparatus and around the nucleus, while the cell cytoplasm was free of fluorescent particles, suggesting that the polyplexes move in the cell toward the nucleus by vesicular transport through the cytoplasm and not by a random diffusion. We found that the plasmids penetrate the cell nucleus without the polymer. Preliminary results in zebra fish and mice demonstrate the potential of ODS to serve as an efficient nonviral vector for in vivo transfection. |
| تدمد: | 1476-5462 0969-7128 |
| DOI: | 10.1038/sj.gt.3302395 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::fd0d5aed4351c60eb19fb234fc77324e https://doi.org/10.1038/sj.gt.3302395 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....fd0d5aed4351c60eb19fb234fc77324e |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14765462 09697128 |
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| DOI: | 10.1038/sj.gt.3302395 |