Role of Intracellular Cationic Liposome–DNA Complex Dissociation in Transfection Mediated by Cationic Lipids
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| Názov: | Role of Intracellular Cationic Liposome–DNA Complex Dissociation in Transfection Mediated by Cationic Lipids |
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| Autori: | Cornelis, S, Vandenbranden, Michel, Ruysschaert, Jean Marie, Elouahabi, Abdelatif |
| Zdroj: | DNA and Cell Biology. 21:91-97 |
| Informácie o vydavateľovi: | Mary Ann Liebert Inc, 2002. |
| Rok vydania: | 2002 |
| Predmety: | 0301 basic medicine, Cytoplasm, Cell Nucleus -- metabolism, Microinjections, Macromolecular Substances, Recombinant Fusion Proteins, Phosphatidylethanolamines -- administration & dosage, Liposomes -- administration & dosage, Amidines, DNA, Recombinant, Phosphatidylethanolamines -- metabolism, Transfection, Amidines -- administration & dosage, Fluorescent Dyes -- analysis, 03 medical and health sciences, Cations -- metabolism, Genes, Reporter, Cations, Chimie, Humans, Reporter, Fluorescent Dyes, Cell Nucleus, Microscopy, Transfection -- methods, 0303 health sciences, Microscopy, Confocal, Amidines -- metabolism, Cations -- administration & dosage, Phosphatidylethanolamines, Biological Transport, DNA, Recombinant -- administration & dosage, Recombinant -- metabolism, beta-Galactosidase, Cytoplasm -- metabolism, Recombinant Fusion Proteins -- biosynthesis, Genes, Liposomes -- metabolism, Lac Operon, Hela Cells, Confocal, Liposomes, beta-Galactosidase -- biosynthesis, HeLa Cells |
| Popis: | The cationic lipid-mediated gene transfer process involves sequential steps: internalization of the cationic lipid-DNA complexes inside the cells via an endocytosis-like mechanism, escape from endosomes, dissociation of the complex, and finally entry of free DNA into the nucleus. However, cationic lipid-DNA complex dissociation in the cytoplasm and the ability of the subsequently released DNA to enter the nucleus have not yet been demonstrated. In this report we showed, using confocal laser scanning analysis, that microinjection of a double fluorescent-labeled cationic lipid-pCMV-LacZ plasmid complex into the cytoplasm of HeLa cells results in efficient complex dissociation. However, the released DNA did not enter the nucleus, and no significant transfection could be detected. In contrast, nuclear microinjection of the cationic lipid-pCMV-LacZ plasmid complex resulted in efficient complex dissociation and transfection of all the cells. Taken together, the data suggest that intracellular dissociation of the cationic lipid-DNA complex is not a limiting step for transfection as previously thought. |
| Druh dokumentu: | Article |
| Popis súboru: | No full-text files |
| Jazyk: | English |
| ISSN: | 1557-7430 1044-5498 |
| DOI: | 10.1089/104454902753604961 |
| Prístupová URL adresa: | https://pubmed.ncbi.nlm.nih.gov/11953008 https://www.ncbi.nlm.nih.gov/pubmed/11953008 https://www.liebertpub.com/doi/abs/10.1089/104454902753604961 https://core.ac.uk/display/8875819 https://online.liebertpub.com/doi/pdf/10.1089/104454902753604961 https://europepmc.org/article/MED/11953008 https://difusion.ulb.ac.be/vufind/Record/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/77596/Details |
| Rights: | Mary Ann Liebert TDM |
| Prístupové číslo: | edsair.doi.dedup.....e03e046ece95414c112083ef3e31abbe |
| Databáza: | OpenAIRE |
Nájsť tento článok vo Web of Science | Abstrakt: | The cationic lipid-mediated gene transfer process involves sequential steps: internalization of the cationic lipid-DNA complexes inside the cells via an endocytosis-like mechanism, escape from endosomes, dissociation of the complex, and finally entry of free DNA into the nucleus. However, cationic lipid-DNA complex dissociation in the cytoplasm and the ability of the subsequently released DNA to enter the nucleus have not yet been demonstrated. In this report we showed, using confocal laser scanning analysis, that microinjection of a double fluorescent-labeled cationic lipid-pCMV-LacZ plasmid complex into the cytoplasm of HeLa cells results in efficient complex dissociation. However, the released DNA did not enter the nucleus, and no significant transfection could be detected. In contrast, nuclear microinjection of the cationic lipid-pCMV-LacZ plasmid complex resulted in efficient complex dissociation and transfection of all the cells. Taken together, the data suggest that intracellular dissociation of the cationic lipid-DNA complex is not a limiting step for transfection as previously thought. |
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| ISSN: | 15577430 10445498 |
| DOI: | 10.1089/104454902753604961 |