Replicating reoviruses with a transgene replacing the codons for the head domain of the viral spike

D. J. M. van den Wollenberg , I. J. C. Dautzenberg , W. Ros , Andrea Lipińska , S. K. van den Hengel , R. C. Hoeben

Abstract

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.
Author D. J. M. van den Wollenberg
D. J. M. van den Wollenberg,,
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, I. J. C. Dautzenberg
I. J. C. Dautzenberg,,
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, W. Ros
W. Ros,,
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, Andrea Lipińska (IFB / IB / LVMB)
Andrea Lipińska,,
- Laboratory of Virus Molecular Biology
, S. K. van den Hengel
S. K. van den Hengel,,
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, R. C. Hoeben
R. C. Hoeben,,
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Journal seriesGene Therapy, ISSN 0969-7128, (A 35 pkt)
Issue year2015
Vol22
No3
Pages267-279
Publication size in sheets0.6
ASJC Classification1311 Genetics; 1312 Molecular Biology; 1313 Molecular Medicine
DOIDOI:10.1038/gt.2014.126
Languageen angielski
Score (nominal)35
Score sourcejournalList
ScoreMinisterial score = 30.0, 05-03-2020, ArticleFromJournal
Ministerial score (2013-2016) = 35.0, 05-03-2020, ArticleFromJournal
Publication indicators WoS Citations = 15; Scopus SNIP (Source Normalised Impact per Paper): 2015 = 0.754; WoS Impact Factor: 2015 = 3.242 (2) - 2015=3.312 (5)
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