Gene transfer to mammalian cells using genetically targeted filamentous bacteriophage.
We have genetically modified filamentous bacteriophage to deliver genes to mammalian cells. In previous studies we showed that noncovalently attached fibroblast growth factor (FGF2) can target bacteriophage to COS-1 cells, resulting in receptor-mediated transduction with a reporter gene. Thus, bacteriophage, which normally lack tropism for mammalian cells, can be adapted for mammalian cell gene transfer. To determine the potential of using phage-mediated gene transfer as a novel display phage screening strategy, we transfected COS-1 cells with phage that were engineered to display FGF2 on their surface coat as a fusion to the minor coat protein, pIII. Immunoblot and ELISA analysis confirmed the presence of FGF2 on the phage coat. Significant transduction was obtained in COS-1 cells with the targeted FGF2-phage compared with the nontargeted parent phage. Specificity was demonstrated by successful inhibition of transduction in the presence of excess free FGF2. Having demonstrated mammalian cell transduction by phage displaying a known gene targeting ligand, it is now feasible to apply phage-mediated transduction as a screen for discovering novel ligands.
Larocca
D
D
Selective Genetics Inc., San Diego, California 92121, USA. laroccad@selectivegenetics.com
Kassner
P D
PD
Witte
A
A
Ladner
R C
RC
Pierce
G F
GF
Baird
A
A
D000818:Animals; D017104:Bacteriophage M13 / Q000235:genetics*; D019556:COS Cells; D036022:Capsid Proteins; D004268:DNA-Binding Proteins / Q000235:genetics; D016222:Fibroblast Growth Factor 2 / Q000235:genetics; D018390:Gene Targeting; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D014760:Viral Fusion Proteins / Q000235:genetics
D036022:Capsid Proteins; D004268:DNA-Binding Proteins; D008164:Luminescent Proteins; D014760:Viral Fusion Proteins; D016222:Fibroblast Growth Factor 2; D049452:Green Fluorescent Proteins