Cargando…
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...
Autores principales: | , , , , , |
---|---|
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 |
_version_ | 1783375662616150016 |
---|---|
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. |
format | Online Article Text |
id | pubmed-6265617 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Portland Press Ltd. |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT hoyoonkhei enhancedtransfectionofamacromolecularligninbaseddnacomplexwithlowcellulartoxicity AT kaidan enhancedtransfectionofamacromolecularligninbaseddnacomplexwithlowcellulartoxicity AT tugeraldinexueen enhancedtransfectionofamacromolecularligninbaseddnacomplexwithlowcellulartoxicity AT deengroshan enhancedtransfectionofamacromolecularligninbaseddnacomplexwithlowcellulartoxicity AT toohengphon enhancedtransfectionofamacromolecularligninbaseddnacomplexwithlowcellulartoxicity AT lohxianjun enhancedtransfectionofamacromolecularligninbaseddnacomplexwithlowcellulartoxicity |