Exploration of peptide motifs for potent non-viral gene delivery highly selective for dividing cells

TY - JOUR

T1 - Exploration of peptide motifs for potent non-viral gene delivery highly selective for dividing cells

AU - Parker, A L

AU - Collins, L

AU - Zhang, X H

AU - Fabre, J W

PY - 2005/12

Y1 - 2005/12

N2 - Background The immunogenicity of viral DNA vectors is an important problem for gene therapy. The use of peptide motifs for gene delivery would largely overcome this problem, and provide a simple, safe and powerful approach for non-viral gene therapy. Methods We explored the functional properties of two motifs: the (Lys)(16) motif (for binding and condensing DNA and probably also nuclear, translocation of plasmids) and the fusogenic peptide motif of influenza virus (for acid-dependent endocytic escape of peptide/DNA particles). The physical properties and gene delivery efficiencies of (Lys)(16)-containing peptides in combination with free fusogenic peptide were evaluated, and compared with a single composite peptide incorporating both moieties. Post-mitotic corneal endothelial cells and growth-arrested HeLa were included, so as not to neglect the question of nuclear translocation of plasmids. Results The fusogenic moiety in the composite peptide was able to adopt an alpha-helical configuration unhindered by the (Lys)(16) moiety, and retained acid-dependent fusogenic properties. The composite peptide gave remarkably high levels of gene delivery to dividing cell lines. However, in marked contrast to (Lys)(16)/DNA complexes plus free fusogenic peptide, the composite peptide was completely ineffective for gene delivery to post-mitotic and growth-arrested cells. Conclusions Attachment of the fusogenic peptide to (Lys)(16) appears to block (Lys)(16)-mediated nuclear translocation of plasmid, but not fusogenic peptide mediated endocytic escape. This strengthens the experimental basis for (Lys)(16)-mediated nuclear translocation of plasmids, and provides a single peptide with potent gene delivery properties, restricted to dividing cells. This property is potentially useful in experimental biology and clinical medicine. Copyright (c) 2005 John Wiley & Sons, Ltd

AB - Background The immunogenicity of viral DNA vectors is an important problem for gene therapy. The use of peptide motifs for gene delivery would largely overcome this problem, and provide a simple, safe and powerful approach for non-viral gene therapy. Methods We explored the functional properties of two motifs: the (Lys)(16) motif (for binding and condensing DNA and probably also nuclear, translocation of plasmids) and the fusogenic peptide motif of influenza virus (for acid-dependent endocytic escape of peptide/DNA particles). The physical properties and gene delivery efficiencies of (Lys)(16)-containing peptides in combination with free fusogenic peptide were evaluated, and compared with a single composite peptide incorporating both moieties. Post-mitotic corneal endothelial cells and growth-arrested HeLa were included, so as not to neglect the question of nuclear translocation of plasmids. Results The fusogenic moiety in the composite peptide was able to adopt an alpha-helical configuration unhindered by the (Lys)(16) moiety, and retained acid-dependent fusogenic properties. The composite peptide gave remarkably high levels of gene delivery to dividing cell lines. However, in marked contrast to (Lys)(16)/DNA complexes plus free fusogenic peptide, the composite peptide was completely ineffective for gene delivery to post-mitotic and growth-arrested cells. Conclusions Attachment of the fusogenic peptide to (Lys)(16) appears to block (Lys)(16)-mediated nuclear translocation of plasmid, but not fusogenic peptide mediated endocytic escape. This strengthens the experimental basis for (Lys)(16)-mediated nuclear translocation of plasmids, and provides a single peptide with potent gene delivery properties, restricted to dividing cells. This property is potentially useful in experimental biology and clinical medicine. Copyright (c) 2005 John Wiley & Sons, Ltd

U2 - 10.1002/jgm.809

DO - 10.1002/jgm.809

M3 - Article

VL - 7

SP - 1545

EP - 1554

JO - JOURNAL OF GENE MEDICINE

JF - JOURNAL OF GENE MEDICINE

IS - 12

ER -