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Enhanced transfection of a macromolecular lignin-based DNA complex with low cellular toxicity

Cationic polymers remain attractive tools for non-viral gene transfer. The effectiveness of these vectors rely on the ability to deliver plasmid DNA (pDNA) into the nucleus of cells. While we have previously demonstrated the potential of Lignin-PGEA-PEGMA as a non-viral gene delivery vector, alterat...

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Autores principales: Ho, Yoon Khei, Kai, Dan, Tu, Geraldine Xue En, Deen, G. Roshan, Too, Heng Phon, Loh, Xian Jun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6265617/
https://www.ncbi.nlm.nih.gov/pubmed/30413612
http://dx.doi.org/10.1042/BSR20181021
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author Ho, Yoon Khei
Kai, Dan
Tu, Geraldine Xue En
Deen, G. Roshan
Too, Heng Phon
Loh, Xian Jun
author_facet Ho, Yoon Khei
Kai, Dan
Tu, Geraldine Xue En
Deen, G. Roshan
Too, Heng Phon
Loh, Xian Jun
author_sort Ho, Yoon Khei
collection PubMed
description Cationic polymers remain attractive tools for non-viral gene transfer. The effectiveness of these vectors rely on the ability to deliver plasmid DNA (pDNA) into the nucleus of cells. While we have previously demonstrated the potential of Lignin-PGEA-PEGMA as a non-viral gene delivery vector, alterations of cellular phenotype and cytotoxicity were observed post transfection. The present study aims to explore transfection conditions for high efficiency and low toxicity of the Lignin-PGEA-PEGMA based gene delivery system. Cellular toxicity was significantly reduced by using the centrifugation protocol, which enables rapid deposition of DNA complexes. Replacement of media post centrifugation resulted in minimal exposure of cells to excess polymers, which were toxic to cells. At an optimized DNA amount (500–750 ng) and molar ratios of nitrogen (N) in polymer to phosphate (P) in pDNA (N/P = 30–40), with the use of a novel transfection enhancer that facilitates endosomal escape and nuclear trafficking, the efficiency of gene delivery was increased significantly 24 h post transfection. The present study demonstrated an appropriately optimized protocol that enabled the utility of a novel cationic polymer blend with a mixture of fusogenic lipids and a histone deacetylate inhibitor in non-viral transfection, thereby providing an attractive alternative to costly commercial gene carriers.
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spelling pubmed-62656172018-12-13 Enhanced transfection of a macromolecular lignin-based DNA complex with low cellular toxicity Ho, Yoon Khei Kai, Dan Tu, Geraldine Xue En Deen, G. Roshan Too, Heng Phon Loh, Xian Jun Biosci Rep Research Articles Cationic polymers remain attractive tools for non-viral gene transfer. The effectiveness of these vectors rely on the ability to deliver plasmid DNA (pDNA) into the nucleus of cells. While we have previously demonstrated the potential of Lignin-PGEA-PEGMA as a non-viral gene delivery vector, alterations of cellular phenotype and cytotoxicity were observed post transfection. The present study aims to explore transfection conditions for high efficiency and low toxicity of the Lignin-PGEA-PEGMA based gene delivery system. Cellular toxicity was significantly reduced by using the centrifugation protocol, which enables rapid deposition of DNA complexes. Replacement of media post centrifugation resulted in minimal exposure of cells to excess polymers, which were toxic to cells. At an optimized DNA amount (500–750 ng) and molar ratios of nitrogen (N) in polymer to phosphate (P) in pDNA (N/P = 30–40), with the use of a novel transfection enhancer that facilitates endosomal escape and nuclear trafficking, the efficiency of gene delivery was increased significantly 24 h post transfection. The present study demonstrated an appropriately optimized protocol that enabled the utility of a novel cationic polymer blend with a mixture of fusogenic lipids and a histone deacetylate inhibitor in non-viral transfection, thereby providing an attractive alternative to costly commercial gene carriers. Portland Press Ltd. 2018-11-30 /pmc/articles/PMC6265617/ /pubmed/30413612 http://dx.doi.org/10.1042/BSR20181021 Text en © 2018 The Author(s). http://creativecommons.org/licenses/by/4.0/This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (http://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Articles
Ho, Yoon Khei
Kai, Dan
Tu, Geraldine Xue En
Deen, G. Roshan
Too, Heng Phon
Loh, Xian Jun
Enhanced transfection of a macromolecular lignin-based DNA complex with low cellular toxicity
title Enhanced transfection of a macromolecular lignin-based DNA complex with low cellular toxicity
title_full Enhanced transfection of a macromolecular lignin-based DNA complex with low cellular toxicity
title_fullStr Enhanced transfection of a macromolecular lignin-based DNA complex with low cellular toxicity
title_full_unstemmed Enhanced transfection of a macromolecular lignin-based DNA complex with low cellular toxicity
title_short Enhanced transfection of a macromolecular lignin-based DNA complex with low cellular toxicity
title_sort enhanced transfection of a macromolecular lignin-based dna complex with low cellular toxicity
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6265617/
https://www.ncbi.nlm.nih.gov/pubmed/30413612
http://dx.doi.org/10.1042/BSR20181021
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