Human-derived NLS enhance the gene transfer efficiency of chitosan |
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Authors: | Diogo B. Bitoque,Joana Morais,Ana V. Oliveira,Raquel L. Sequeira,Sofia M. Calado,Tiago M. Fortunato,Só nia Simã o,Ana M. Rosa da Costa,Gabriela A. Silva |
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Affiliation: | 1.CEDOC, NOVA Medical School, Universidade Nova de Lisboa, Campo dos Mártires da Pátria, 130, 1169-056 Lisboa, Portugal;2.Algarve Chemistry Research Centre (CIQA), University of Algarve, Faro, Portugal;3.Centre for Biomedical Research (CBMR), University of Algarve, Campus Gambelas, 8005 Faro, Portugal |
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Abstract: | Nuclear import is considered as one of the major limitations for non-viral gene delivery systems and the incorporation of nuclear localization signals (NLS) that mediate nuclear intake can be used as a strategy to enhance internalization of exogenous DNA.In this work, human-derived endogenous NLS peptides based on insulin growth factor binding proteins (IGFBP), namely IGFBP-3 and IGFBP-5, were tested for their ability to improve nuclear translocation of genetic material by non-viral vectors. Several strategies were tested to determine their effect on chitosan mediated transfection efficiency: co-administration with polyplexes, co-complexation at the time of polyplex formation, and covalent ligation to chitosan. Our results show that co-complexation and covalent ligation of the NLS peptide derived from IGFBP-3 to chitosan polyplexes yields a 2-fold increase in transfection efficiency, which was not observed for NLS peptide derived from IGFBP-5.These results indicate that the integration of IGFBP-NLS-3 peptides into polyplexes has potential as a strategy to enhance the efficiency of non-viral vectors. |
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Keywords: | chitosan gene therapy HEK293T cells IGFBP nuclear localization signals |
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