Human-derived NLS enhance the gene transfer efficiency of chitosan

Biosci Rep. 2021 Jan 29;41(1):BSR20201026. doi: 10.1042/BSR20201026.

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.

Keywords: HEK293T cells; IGFBP; chitosan; gene therapy; nuclear localization signals.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chitosan / metabolism*
  • Gene Transfer Techniques / standards*
  • HEK293 Cells
  • Humans
  • Insulin-Like Growth Factor Binding Protein 3 / metabolism
  • Insulin-Like Growth Factor Binding Protein 5 / metabolism
  • Nuclear Localization Signals*

Substances

  • Insulin-Like Growth Factor Binding Protein 3
  • Insulin-Like Growth Factor Binding Protein 5
  • Nuclear Localization Signals
  • Chitosan