title: Gene transfer to mammalian cells using genetically targeted filamentous bacteriophage.
abstract: 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.
authors: Selective Genetics Inc., San Diego, California 92121, USA. laroccad@selectivegenetics.com
authors: D
authors: D
authors: Larocca
authors: P D
authors: PD
authors: Kassner
authors: A
authors: A
authors: Witte
authors: R C
authors: RC
authors: Ladner
authors: G F
authors: GF
authors: Pierce
authors: A
authors: A
authors: Baird
mesh_terms: 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
chemical_list: D036022:Capsid Proteins; D004268:DNA-Binding Proteins; D008164:Luminescent Proteins; D014760:Viral Fusion Proteins; D016222:Fibroblast Growth Factor 2; D049452:Green Fluorescent Proteins