Cell-type specific targeting and gene expression using a variant of polyoma VP1 virus-like particles.
The variant VP1-Z of the polyomavirus coat protein VP1 has been recently described as an engineered fusion protein of VP1 and the antibody binding domain protein Z. This construct is able to specifically bind and functionally present antibodies on the surface of virus-like particles of VP1-Z. Here we demonstrate that with the binding of Herceptin, an antibody directed against the receptor tyrosine kinase ErbB2, a cell type-specific targeting was established. ErbB2-positive cell lines were transduced with different plasmids encoding eGFP or beta-galactosidase. With both reporter systems functional gene expression in transduced cells could be observed. The transduction was strictly dependent on the use of a ternary complex formed of VLPs of VP1-Z, Herceptin, and the reporter plasmid DNA. The use of single components or ErbB2-negative cell lines did not result in functional gene transfer. The transduction was also completely dependent on the use of chloroquine, a lysosomotropic reagent. This indicates that the complex is internalized by ErbB2-mediated endocytosis.
Gleiter
Stefan
S
Martin-Luther-Universität Halle, Institut für Biotechnologie, Kurt-Mothes Strasse 3, D-06120 Halle, Germany.
Lilie
Hauke
H
D000911:Antibodies, Monoclonal / Q000378:metabolism; D061067:Antibodies, Monoclonal, Humanized; D001943:Breast Neoplasms / Q000378:metabolism; D036022:Capsid Proteins / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002738:Chloroquine / Q000494:pharmacology; D015967:Gene Expression Regulation, Viral / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D014644:Genetic Variation / Q000235:genetics; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011120:Polyomavirus / Q000737:chemistry; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000096:biosynthesis / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*; D000068878:Trastuzumab; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D036022:Capsid Proteins; D002738:Chloroquine; D008164:Luminescent Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; D001616:beta-Galactosidase; C062481:VP1 protein, polyomavirus