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Publicação
Human-derived NLS enhance the gene transfer efficiency of chitosan
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Orientador(es)
Resumo(s)
Nuclear import is considered 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.
Descrição
The authors acknowledge Dr. Jean Bennett (University of Pennsylvania, USA) for kindly providing the GFP plasmid and Dr. Lyn Schedlich and Sue Firth for kindly providing the NLS plasmids. The authors also acknowledge the financial support of Fundação para a Ciência e Tecnologia (PTDC/SAL -BEB/098475/2008 to Gabriela A. Silva, PD/BD/52424/2013 individual fellowship to Diogo Bitoque, SFRH/BD/70318/2010 individual fellowship to Ana V. Oliveira, SFRH/BD/76873/2011 individual fellowship to Sofia Calado, SFRH/BPD/78404/2011 individual fellowship to Sónia Simão, IBB/LA under the project PEst- OE/EQB/LA0023/2013, and PEst-OE_QUI_UI4023_2011) and the Marie Curie Reintegration Grant (PIRG-GA-2009-249314 to Gabriela A. Silva) under the FP7 program. iNOVA4Health - UID/Multi/04462/2013, a program financially supported by Fundação para a Ciência e Tecnologia / Ministério da Educação e Ciência, through national funds and co-funded by FEDER under the PT2020 Partnership Agreement is also acknowledged.
Palavras-chave
gene therapy chitosan nuclear localization signals IGFBP HEK293T cells Biotechnology