CEDOC, NOVA Medical School, Universidade Nova de Lisboa, Campo dos Mártires da Pátria, 130, 1169-056 Lisboa, Portugal.
Algarve Chemistry Research Centre (CIQA), University of Algarve, Faro, Portugal.
Biosci Rep. 2021 Jan 29;41(1). doi: 10.1042/BSR20201026.
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.
核内输入被认为是非病毒基因传递系统的主要限制因素之一，而介导核摄取的核定位信号（NLS）的掺入可以用作增强外源性 DNA 内化的策略。在这项工作中，基于胰岛素生长因子结合蛋白（IGFBP）的人源性内源性 NLS 肽，即 IGFBP-3 和 IGFBP-5，被测试其通过非病毒载体改善遗传物质核易位的能力。测试了几种策略来确定它们对壳聚糖介导的转染效率的影响：与多聚物共给药、在多聚物形成时共复合以及与壳聚糖共价连接。我们的结果表明，将 IGFBP-3 衍生的 NLS 肽与壳聚糖多聚物共复合和共价连接可使转染效率提高 2 倍，而 IGFBP-5 衍生的 NLS 肽则没有观察到这种情况。这些结果表明，将 IGFBP-NLS-3 肽整合到多聚物中具有作为增强非病毒载体效率的策略的潜力。
