Targeting of High-Capacity Adenoviral Vectors
| Autor: | Gudrun Schiedner, Andreas Herrmann, Steffi Hussmann, Stefan Kochanek, Christoph Volpers, Volker Biermann, Helmut Kewes, Anke Stock |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Coxsackie and Adenovirus Receptor-Like Membrane Protein
viruses Blotting Western Genetic Vectors Protozoan Proteins Genome Viral Biology Ligands medicine.disease_cause Recombinant virus Binding Competitive Epitope Virus Adenoviridae Epitopes Transduction (genetics) Plasmid Transduction Genetic Tumor Cells Cultured Genetics medicine Humans Endothelium Cloning Molecular Molecular Biology Cells Cultured Models Genetic Genetic transfer Gene Transfer Techniques Muscle Smooth beta-Galactosidase Molecular biology Protein Structure Tertiary Helper virus Receptors Virus Molecular Medicine Peptides Helper Viruses Oligopeptides Plasmids |
| Zdroj: | Human Gene Therapy. 12:1757-1769 |
| ISSN: | 1557-7422 1043-0342 |
| DOI: | 10.1089/104303401750476258 |
| Popis: | High-capacity adenoviral (HC-Ad) vectors contain only the noncoding termini of the viral genome, can deliver large DNA fragments of up to 36 kb into target cells, and feature reduced toxicity and prolonged transgene expression in vivo. To enhance the potential of HC-Ad vectors to transduce specific cell types, we constructed a versatile infectious new helper virus plasmid that can be used readily to introduce peptide ligands into the HI loop of the fiber knob domain of Ad5-based HC-Ad vectors. Helper viruses with a 6x-His epitope or Arg-Gly-Asp (RGD) peptide insertion retained the full infectivity of the wild-type helper virus. The RGD-modified helper virus was used for production of a capsid-modified HC-Ad vector expressing beta-galactosidase. The RGD HC-Ad vector transduced the ovarian carcinoma cell lines SK-OV-3 and OVCAR-3 with 4- to 20-fold higher efficiency, compared to unmodified vectors. Transduction of both primary vascular smooth muscle cells as well as primary human endothelial cells was increased up to 15-fold with the RGD-modified vector. Competition experiments with recombinant knob protein and different RGD peptides indicated that the RGD-mediated transduction was Coxsackie and Adenovirus receptor (CAR)-independent and involved integrin alpha(v)beta(5). The use of fiber-modified helper viruses in the last amplification step of HC-Ad vector production allows for convenient and efficient targeting of these vectors towards different cell types. Targeting strategies will increase the spectrum of applications for HC-Ad vectors and will further add to their safety. |
| Databáze: | OpenAIRE |
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