Cardiff University | Prifysgol Caerdydd ORCA
Online Research @ Cardiff 
WelshClear Cookie - decide language by browser settings

Nuclear localisation and pDNA condensation in non-viral gene delivery

Collins, Elizabeth, Birchall, James Caradoc ORCID:, Williams, Julie L. and Gumbleton, Mark ORCID: 2007. Nuclear localisation and pDNA condensation in non-viral gene delivery. The Journal of Gene Medicine 9 (4) , pp. 265-274. 10.1002/jgm.1015

Full text not available from this repository.


Background Non-viral gene delivery vectors are multi-component systems reflecting various functionalities required for effective cell transfection, including DNA condensation, promotion of cell membrane interactions and provision for subcellular targeting through endosomal escape and/or nuclear delivery. Elements mediating these functions will clearly display inter-dependency. In this study we sought to explore the relationship within non-viral vectors of condensation and nuclear localisation. Methods Binary, tertiary and quaternary vectors were prepared with combinations of pDNA, DOTAP lipid, the polycation peptide protamine and either SV40 nuclear localisation sequence peptide (‘SV40 NLS’) or a one amino acid substituted mutant of SV40 NLS (‘mutant sequence’). The efficiency of pDNA condensation was determined by gel electrophoresis and quantitative fluorescence spectroscopy. Transfection efficiency was examined in mammalian cells in vitro using standard methods, by electroporation to bypass the plasma membrane barrier and in cells arrested in G0/G1 cell cycle phase to examine the effect of cell division and nuclear membrane disruption. Results Small NLS peptide sequences, despite possessing a significant proportion of basic amino acids, display minimal pDNA-condensing ability when compared to larger polycations such as protamine. In standard in vitro cell adherent transfection studies the predominant elements affording enhanced gene expression were effective pDNA condensation and lipid enhancement of cell membrane interactions. These features conversely hinder efficient gene expression in cells that have undergone electroporation. The benefit of SV40 NLS was only apparent when used in non-dividing cell populations. Conclusions Whilst effective levels of non-viral-mediated gene expression generally rely on efficient condensation of pDNA and enhanced interactions with cellular membranes, non-covalently associated NLS within a multi-component non-viral gene vector appears to contribute benefit in sustaining gene expression in non-dividing cells. Copyright © 2007 John Wiley & Sons, Ltd.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Pharmacy
Subjects: R Medicine > R Medicine (General)
Uncontrolled Keywords: Nuclear localisation ; pDNA condensation ; Non-vira ; Gene delivery ; Electroporation ; Cell cycle
ISSN: 1521-2254
Last Modified: 17 Oct 2022 08:45

Citation Data

Cited 32 times in Scopus. View in Scopus. Powered By Scopus® Data

Actions (repository staff only)

Edit Item Edit Item