Replicating reoviruses with a transgene replacing the codons for the head domain of the viral spike.
| Autor: | van den Wollenberg DJ; Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands., Dautzenberg IJ; Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands., Ros W; Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands., Lipińska AD; Department of Virus Molecular Biology, Intercollegiate Faculty of Biotechnology UG-MUG, University of Gdańsk, Gdańsk, Poland., van den Hengel SK; Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands., Hoeben RC; Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands. |
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| Jazyk: | angličtina |
| Zdroj: | Gene therapy [Gene Ther] 2015 Mar; Vol. 22 (3), pp. 267-79. Date of Electronic Publication: 2015 Jan 15. |
| DOI: | 10.1038/gt.2014.126 |
| Abstrakt: | The capacity to modify the reovirus genome facilitates generation of new therapeutic reoviruses. We describe a method for generating replication-competent reoviruses carrying a heterologous transgene. The strategy is based on the expanded-tropism reovirus mutant jin-3, which can infect cells independent of the reovirus receptor junction-adhesion molecule A (JAM-A). Jin-3 harbors a mutation in the S1 segment, resulting in a G196R substitution in the tail of the spike protein σ1. The use of the jin-3 tail-encoding S1 segment allows replacing the codons for the JAM-A-binding head domain by up to 522 nucleotides of foreign sequences, without exceeding the size of the wild-type S1 segment. We inserted the codons for the porcine teschovirus-1 2A element fused with those encoding the fluorescent protein iLOV. Replicating rS1His-2A-iLOV reoviruses were generated by co-transfection of expression plasmids for all reovirus segments. These reoviruses contain the S1His-2A-iLOV segment in the absence of the wild-type S1 segment. Density-gradient centrifugation confirmed the association of the σ1-tail fragment with the capsid. Both JAM-A-positive and -negative cells exposed to the rS1His-2A-iLOV reoviruses exhibited iLOV fluorescence, confirming the jin-3-derived expanded-tropism phenotype. These data demonstrated the feasibility of generating decapitated replication-competent T3D reoviruses carrying a heterologous transgene. |
| Databáze: | MEDLINE |
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