Binding properties, cell delivery, and gene transfer of adenoviral penton base displaying bacteriophage.
The penton base of adenovirus mediates viral attachment to integrin receptors and particle internalisation, properties that can be exploited to reengineer prokaryotic viruses for the infection of mammalian cells. We report that filamentous phage displaying either the full-length penton base gene or a central region of 107 amino acids on their surface were able to bind, internalise, and transduce mammalian cells expressing integrin receptors. Both phage bound alphavbeta3, alphavbeta5, alpha3beta1, and alpha5beta1 integrin subtypes. Cell-binding was shown by electron microscopy; internalisation was investigated by immunofluorescence and confirmed by micropanning. As it has been described for adenovirus, pharmacologic disruption of phosphoinositide-30H kinase, but not of myosin light-chain kinase, inhibited phage internalisation. Recombinant phage encoding an eukaryotic expression cassette was able to mediate gene expression in mammalian cells. Taken together, these data open insights for the exploit of recombinant phage for integrin-targeted gene delivery.
Department of Genetics and Molecular Biology, University La Sapienza, Rome, Italy.
M
M
Di Giovine
B
B
Salone
Y
Y
Martina
V
V
Amati
G
G
Zambruno
E
E
Cundari
C M
CM
Failla
I
I
Saggio
D000256:Adenoviridae / Q000235:genetics*; D001435:Bacteriophages / Q000235:genetics*; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D002352:Carrier Proteins / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D016023:Integrins / Q000378:metabolism; D008854:Microscopy, Electron; D009842:Oligopeptides / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D014161:Transduction, Genetic
D036022:Capsid Proteins; D002352:Carrier Proteins; D016023:Integrins; D009842:Oligopeptides; C094697:glutamyl-isoleucyl-leucyl-aspartyl-valine; C043322:penton protein, adenovirus; D019869:Phosphatidylinositol 3-Kinases