An RGD-oligolysine peptide: a prototype construct for integrin-mediated gene delivery.
We have synthesized a linear, bifunctional peptide that comprises an integrin-targeting domain containing an arginine-glycine-aspartic acid tripeptide motif and a DNA-binding moiety consisting of a short stretch of 16 lysine residues. This peptide can form distinctive, condensed complexes with DNA and is capable of mediating its delivery and expression in a variety of mammalian cells in culture. Internalization is mediated by cell surface integrin receptors via a mechanism that is known to be phagocytic. We have analyzed the relationship between DNA and peptide and have investigated the conditions suitable for optimal gene delivery. The formation of condensed peptide DNA complexes leads to resistance to nuclease degradation. The level of reporter gene expression obtained is dependent on the peptide-to-DNA ratio and is enhanced in the presence of the endosomal buffer chloroquine, polyethyleneimine, and deactivated adenovirus during gene delivery. Under optimal conditions the levels of reporter gene expression obtained approach or even exceed those obtained with DNA delivered with the commercial liposome Lipofectamine. The ability to produce an efficient gene delivery system using small, easily modified, and well-defined constructs that have no constraint of particle size demonstrates the advantages of integrin-targeting peptides for gene transfer.
Harbottle
R P
RP
Division of Biomedical Sciences, Molecular Genetics, Imperial College School of Medicine at St. Mary's, London, UK.
Cooper
R G
RG
Hart
S L
SL
Ladhoff
A
A
McKay
T
T
Knight
A M
AM
Wagner
E
E
Miller
A D
AD
Coutelle
C
C
D016475:3T3 Cells; D000818:Animals; D018938:Caco-2 Cells; D002738:Chloroquine; D004268:DNA-Binding Proteins / Q000138:chemical synthesis / Q000378:metabolism; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D016023:Integrins / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics; D051379:Mice; D009842:Oligopeptides / Q000138:chemical synthesis / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011094:Polyethyleneimine; D011107:Polylysine / Q000138:chemical synthesis / Q000378:metabolism; D014162:Transfection
D004268:DNA-Binding Proteins; D016023:Integrins; D009842:Oligopeptides; D011107:Polylysine; C047981:arginyl-glycyl-aspartic acid; D002738:Chloroquine; D011094:Polyethyleneimine; D008156:Luciferases