Targeted gene transfer system using a streptavidin-transforming growth factor-alpha chimeric protein.
The previously reported streptavidin-TGFalpha chimeric protein-based delivery system (Ohno and Meruelo, DNA Cell Biol. 15:401-406, 1996) could efficiently transfer protein molecules into A431 cells via the epidermal growth factor (EGF) receptor. We have modified this delivery system for the transfer of DNA. For this purpose, we have linked the chimeric protein ST-TGFalpha to DNA through biotinylated polylysine molecules. We show with this system, in the presence of the endosome-destabilizing reagent chloroquine, an average of 50-fold increase in reporter gene expression in comparison with polylysine DNA complexes alone. This gene expression is specific for EGF receptor-expressing cells and is blocked by EGF-binding molecules. These results suggest that the ST-TGFalpha biotinylated polylysine system could be used to deliver DNA to targeted cells.
Garcia-Espana
A
A
Department of Pathology and Kaplan Cancer Center, New York University Medical Center, NY 10016, USA.
Biria
S
S
Malumbres
M
M
Levin
B
B
Meruelo
D
D
Pellicer
A
A
D000818:Animals; D019921:Biotinylation; D019556:COS Cells; D002294:Carcinoma, Squamous Cell / Q000473:pathology; D002461:Cell Line, Transformed; D002522:Chlorocebus aethiops; D002738:Chloroquine / Q000494:pharmacology; D004274:DNA, Recombinant / Q000008:administration & dosage; D004337:Drug Carriers; D004705:Endocytosis; D011992:Endosomes / Q000187:drug effects; D066246:ErbB Receptors / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects*; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D009363:Neoplasm Proteins / Q000378:metabolism*; D011107:Polylysine / Q000008:administration & dosage; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology*; D013539:Simian virus 40; D019809:Streptavidin / Q000235:genetics / Q000494:pharmacology*; D014162:Transfection / Q000379:methods*; D016211:Transforming Growth Factor alpha / Q000235:genetics / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D004274:DNA, Recombinant; D004337:Drug Carriers; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; D016211:Transforming Growth Factor alpha; D011107:Polylysine; D002738:Chloroquine; D019809:Streptavidin; D008156:Luciferases; D066246:ErbB Receptors