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Full-Text Articles in Dentistry
Correlation Between The Levels Of Survivin And Survivin Promoter-Driven Gene Expression In Cancer And Non-Cancer Cells, Krystyna Konopka, Christopher Spain, Allison Yen, Nathan Overlid, Senait Gebremedhin, Nejat Düzgüneş
Correlation Between The Levels Of Survivin And Survivin Promoter-Driven Gene Expression In Cancer And Non-Cancer Cells, Krystyna Konopka, Christopher Spain, Allison Yen, Nathan Overlid, Senait Gebremedhin, Nejat Düzgüneş
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Survivin, a member of the inhibitor of apoptosis (IAP) protein family, is associated with malignant transformation and is over-expressed in most human tumors. Using lipoplex-mediated transfection, we evaluated the activity of the reporter enzyme, luciferase, expressed from plasmids encoding the enzyme under the control of either the cytomegalovirus (CMV) or survivin promoters, in tumor- and non-tumor-derived human and murine cells. We also examined whether there is a correlation between the survivin promoter-driven expression of luciferase and the level of endogenous survivin. Human cancer cells (HeLa, KB, HSC-3, H357, H376, H413), oral keratinocytes, GMSM-K, and chemically immortalized human mammary cells, 184A-1, …
Serum Decreases The Size Of Metafectene- And Genejammer-Dna Complexes But Does Not Affect Significantly Their Transfection Activity In Sccvii Murine Squamous Cell Carcinoma Cells, Krystyna Konopka, Nathan Overlid, Anitha C. Nagaraj, Nejat Düzgüneş
Serum Decreases The Size Of Metafectene- And Genejammer-Dna Complexes But Does Not Affect Significantly Their Transfection Activity In Sccvii Murine Squamous Cell Carcinoma Cells, Krystyna Konopka, Nathan Overlid, Anitha C. Nagaraj, Nejat Düzgüneş
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Cationic liposome-DNA (lipoplexes) or polymer-DNA (polyplexes) complexes have been used to deliver therapeutic genes, both in vitro and in vivo. However, gene transfer by these non-viral vectors is usually inhibited by biological milieu. A relatively high efficiency of transfection could be achieved in human oral cancer cells transfected with the polycationic liposome, Metafectene, and the polyamine reagent, GeneJammer, in the presence of 60% fetal bovine serum (FBS) (Konopka et al., Cell. Mol. Biol. Lett. 10 (2005) 455-470). Here, we examined the efficacy of these vectors to deliver β-galactosidase (β-gal), luciferase and Herpes Simplex Virus thymidine kinase (HSV-tk) genes to SCCVII …
Efficient Gene Transfer By Transferrin Lipoplexes In The Presence Of Serum, Conchita Tros De Ilarduya, Nejat Düzgüneş
Efficient Gene Transfer By Transferrin Lipoplexes In The Presence Of Serum, Conchita Tros De Ilarduya, Nejat Düzgüneş
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Cationic lipids are being used increasingly as reagents for gene delivery both in vitro and in vivo. One of the limitations to the application of cationic lipid-DNA complexes (lipoplexes) in vivo is the inhibition of gene delivery by serum. In this study, we have shown that transferrin (Tf)-lipoplexes, which had transferrin adsorbed at their surface via electrostatic interactions, are much more effective than plain lipoplexes in transfecting cells in the presence of relatively high concentrations (up to 60%) of fetal bovine serum (FBS). Serum even enhanced transfection by Tf-lipoplexes composed of 1,2-dioleoyl-3-(trimethylammonium) propane (DOTAP)/dioleoylphosphatidylethanolamine (DOPE)/pCMVLacZ at high lipid/DNA (+/-) charge …
Mechanisms Of Gene Transfer Mediated By Lipoplexes Associated With Targeting Ligands Or Ph-Sensitive Peptides, Sérgio Simões, Vladimir Slepushkin, Pedro Pires, Rogério Gaspar, Maria C. Pedroso De Lima, Nejat Düzgüneş
Mechanisms Of Gene Transfer Mediated By Lipoplexes Associated With Targeting Ligands Or Ph-Sensitive Peptides, Sérgio Simões, Vladimir Slepushkin, Pedro Pires, Rogério Gaspar, Maria C. Pedroso De Lima, Nejat Düzgüneş
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Association of a targeting ligand such as transferrin, or an endosome disrupting peptide such as GALA, with cationic liposome-DNA complexes ('lipoplexes') results in a significant enhancement of transfection of several cell types. Although these strategies can overcome some of the barriers to gene delivery by lipoplexes, the mechanisms by which they actually enhance transfection is not known. In studies designed to establish the targeting specificity of transferrin, we found that apo-transferrin enhances transfection to the same extent as transferrin, indicating that internalization of the lipoplexes is mostly independent of transferrin receptors. These observations were reinforced by results obtained from competitive …
Interaction Of Cationic Liposomes And Their Dna Complexes With Monocytic Leukemia Cells, Pedro Pires, Sérgio Simões, Shlomo Nir, Rogério Gaspar, Nejat Düzgüneş, Maria C. Pedroso De Lima
Interaction Of Cationic Liposomes And Their Dna Complexes With Monocytic Leukemia Cells, Pedro Pires, Sérgio Simões, Shlomo Nir, Rogério Gaspar, Nejat Düzgüneş, Maria C. Pedroso De Lima
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Cationic liposomes complexed with DNA have been used extensively as non-viral vectors for the intracellular delivery of reporter or therapeutic genes in culture and in vivo. We examined the relationship between the characteristics of the lipoplexes, their mode of interaction with monocytic THP-1 cells and their ability to transfect these cells. We determined the size and ζ potential of cationic liposomes (composed of 1,2-dioleoyl-3-(trimethylammonium) propane (DOTAP) and its mixtures with neutral lipids), and lipoplexes at different (+/-) charge ratios. As the (+/-) charge ratio of the lipoplexes decreased to (1/1), a significant reduction in ζ potential and an increase in …
Cationic Liposome-Mediated Expression Of Hiv-Regulated Luciferase And Diphtheria Toxin A Genes In Hela Cells Infected With Or Expressing Hiv, Krystyna Konopka, Gail S. Harrison, Philip L. Felgner, Nejat Düzgüneş
Cationic Liposome-Mediated Expression Of Hiv-Regulated Luciferase And Diphtheria Toxin A Genes In Hela Cells Infected With Or Expressing Hiv, Krystyna Konopka, Gail S. Harrison, Philip L. Felgner, Nejat Düzgüneş
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HIV-regulated expression of the diphtheria toxin A fragment gene (HIV-DT-A) is a potential gene therapy approach to AIDS. Since cationic liposomes are safe and non-immunogenic for in vivo gene delivery, we examined whether LipofectAMINE or DMRIE reagent could mediate the transfection of HIV-DT-A (pTHA43) or the HIV-regulated luciferase gene (pLUCA43) into HIV-infected or uninfected HeLa cells. pLUCA43 was expressed at a 103-fold higher level in HeLa/LAV cells than in uninfected HeLa cells, while the extent of expression of RSV-regulated luciferase was the same in both cell lines. Co-transfection of HeLa cells with pTHA43 and the proviral HIV clone, HXBΔBgl, resulted …