Supramolecular structure and nuclear targeting efficiency determine the enhancement of transfection by modified polylysines

C. K. Chan, T. Senden, D. A. Jans

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69 Citations (Scopus)

Abstract

Polylysine (ply) has been used as a DNA carrier in nonviral gene delivery systems because it forms complexes with plasmid DNA via charge interaction, and condenses it into a compact structure. We have recently shown that cross-linking nuclear localization sequences (NLSs) to ply can enhance transfection by conferring specific recognition by the cellular nuclear import 'receptor', the NLS-binding importin α/β heterodimer. The present study examines and correlates for the first time the effect of the lysine/nucleotide (Ly/Nu) ratio on transfection, recognition by importin α/β, and structure as determined using electron microscopy (EM) and atomic force microscopy (AFM), for ply-DNA complexes with and without NLSs or mutant versions thereof. Intriguingly, we observed two distinct peaks of transfection enhancement at Ly/Nu ratios of 0.4 and 4.0, attributable to specific NLS recognition by importins and DNA compaction, respectively. The results indicate a clear correlation between the ply-DNA structure, importin α/β recognition, and gene transfer efficiency, thus underlining the importance of using ply-DNA at the optimal Ly/Nu ratio.

Original languageEnglish
Pages (from-to)1690-1697
Number of pages8
JournalGene Therapy
Volume7
Issue number19
DOIs
Publication statusPublished - 1 Jan 2000
Externally publishedYes

Keywords

  • Atomic force microscopy
  • Electron microscopy
  • Importin subunits
  • Nonviral DNA transfer
  • Nuclear localization signals

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