Manipulation of both virus- and cell-specific factors is required for robust transient replication of a hepatitis C virus genotype 3a sub-genomic replicon Article Swipe
Lorna Kelly
,
Anjna Badhan
,
Grace Roberts
,
Jean L. Mbisa
,
Mark Harris
·
YOU?
·
· 2017
· Open Access
·
· DOI: https://doi.org/10.1099/jgv.0.000932
YOU?
·
· 2017
· Open Access
·
· DOI: https://doi.org/10.1099/jgv.0.000932
Hepatitis C virus (HCV) genotype (GT) 3 is the second most prevalent of the seven HCV genotypes and exhibits the greatest resistance to the highly potent, direct-acting antivirals (DAAs) that are currently in use. Previously a stable cell line harbouring the S52 GT3 sub-genomic replicon (SGR) was established, but this SGR was unable to robustly replicate transiently. As transient SGRs are a critical tool in the development of DAAs, and in the study of viral resistance, we sought to establish a transient SGR system based on S52. Next-generation sequencing was used to identify putative culture-adaptive substitutions that had arisen during long-term selection of the S52 SGR. A subset of these substitutions was built back into the S52 SGR in the context of a CpG/UpA-low luciferase reporter, with a single point mutation in NS4A conferring the greatest replication capability upon S52. Modification of the innate immune-sensing pathways of Huh7.5 hepatoma cells by expression of the parainfluenza virus type 5 V protein and SEC14L2 resulted in a further enhancement of S52 replication. Furthermore, this transiently replicating SGR showed genotype-specific differences in sensitivity to two clinically relevant NS5A DAAs. In conclusion, we report that a single substitution in NS4A, coupled with host cell modifications, enabled robust levels of transient replication by the GT3 S52 SGR. This system will have beneficial uses in both basic research into the unique aspects of GT3 biology and drug discovery.
Related Topics
Concepts
Biology
NS5A
Virology
Replicon
Genotype
Viral replication
Virus
Hepatitis C virus
Context (archaeology)
Genetics
Hepacivirus
Gene
Genome
Paleontology
Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1099/jgv.0.000932
- OA Status
- hybrid
- Cited By
- 5
- References
- 37
- Related Works
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- OpenAlex ID
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All OpenAlex metadata
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https://openalex.org/W2757637661Canonical identifier for this work in OpenAlex
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https://doi.org/10.1099/jgv.0.000932Digital Object Identifier
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Manipulation of both virus- and cell-specific factors is required for robust transient replication of a hepatitis C virus genotype 3a sub-genomic repliconWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
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2017Year of publication
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2017-10-01Full publication date if available
- Authors
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Lorna Kelly, Anjna Badhan, Grace Roberts, Jean L. Mbisa, Mark HarrisList of authors in order
- Landing page
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https://doi.org/10.1099/jgv.0.000932Publisher landing page
- Open access
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YesWhether a free full text is available
- OA status
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hybridOpen access status per OpenAlex
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https://doi.org/10.1099/jgv.0.000932Direct OA link when available
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Biology, NS5A, Virology, Replicon, Genotype, Viral replication, Virus, Hepatitis C virus, Context (archaeology), Genetics, Hepacivirus, Gene, Genome, PaleontologyTop concepts (fields/topics) attached by OpenAlex
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5Total citation count in OpenAlex
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2023: 1, 2022: 1, 2020: 2, 2018: 1Per-year citation counts (last 5 years)
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10Other works algorithmically related by OpenAlex
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