A strategy for genetic modification of the spike-encoding segment of human reovirus T3D for reovirus targeting
| Autor: | Iris J. C. Dautzenberg, S K van den Hengel, Onno Kranenburg, Rob C. Hoeben, D J M van den Wollenberg, Steve J. Cramer |
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| Rok vydání: | 2008 |
| Předmět: |
Junctional Adhesion Molecules
Cell Survival viruses Genetic enhancement Molecular Sequence Data Reoviridae Gene delivery Ligands Transduction (genetics) Transduction Genetic Cell Line Tumor Neoplasms Genetics Humans Amino Acid Sequence Cloning Molecular Immunoglobulin Fragments Mammalian orthoreovirus 3 Molecular Biology Orthoreovirus Oncolytic Virotherapy biology virus diseases Gene targeting Bystander Effect Genetic Therapy biology.organism_classification Virology Oncolytic virus Oncolytic Viruses Capsid Gene Targeting Molecular Medicine Genetic Engineering Cell Adhesion Molecules Sequence Alignment |
| Zdroj: | Gene Therapy. 15:1567-1578 |
| ISSN: | 1476-5462 0969-7128 |
| DOI: | 10.1038/gt.2008.118 |
| Popis: | Human Orthoreovirus Type 3 Dearing is not pathogenic to humans and has been evaluated clinically as an oncolytic agent. Its transduction efficiency and the tumor cell selectivity may be enhanced by incorporating ligands for alternative receptors. However, the genetic modification of reoviruses has been difficult, and genetic targeting of reoviruses has not been reported so far. Here we describe a technique for generating genetically targeted reoviruses. The propagation of wild-type reoviruses on cells expressing a modified sigma 1-encoding segment embedded in a conventional RNA polymerase II transcript leads to substitution of the wild-type genome segment by the modified version. This technique was used for generating reoviruses that are genetically targeted to an artificial receptor expressed on U118MG cells. These cells lack the junction adhesion molecule-1 and therefore resist infection by wild-type reoviruses. The targeted reoviruses were engineered to carry the ligand for this receptor at the C terminus of the sigma 1 spike protein. This demonstrates that the C terminus of the sigma 1 protein is a suitable locale for the insertion of oligopeptide ligands and that targeting of reoviruses is feasible. The genetically targeted viruses can be propagated using the modified U118MG cells as helper cells. This technique may be applicable for the improvement of human reoviruses as oncolytic agents. |
| Databáze: | OpenAIRE |
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