ArticleViruses2019
Reverse Genetics of RNA Viruses: ISA-Based Approach to Control Viral Population Diversity without Modifying Virus Phenotype.
Article in Viruses, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 16 citations in OpenAlex.
- West Nile virus vaccine candidates attenuated by dinucleotide enrichment are immunogenic and protective against lethal infection.PLoS pathogens · 2025Article
- Development of GFP-expressing infectious clones for PRRSV using TAR cloning for antiviral drug screening.Npj viruses · 2025Article
- Infectious Subgenomic Amplicon Strategies for Japanese Encephalitis and West Nile Viruses.Journal of medical virology · 2025Article
- Article
- Rescue and in vitro characterization of a divergent TBEV-Eu strain from the Netherlands.Scientific reports · 2023Article
- Development and characterization of chimera of yellow fever virus vaccine strain and Tick-Borne encephalitis virus.PloS one · 2023Article
- Development of a rapid reverse genetics system for feline coronavirus based on TAR cloning in yeast.Frontiers in microbiology · 2023Article
- Different viral genes modulate virulence in model mammal hosts andFrontiers in microbiology · 2023Article
- Article
- Rapid reconstruction of porcine reproductive and respiratory syndrome virus using synthetic DNA fragments.Computational and structural biotechnology journal · 2021Article
- Molecular Determinants of West Nile Virus Virulence and Pathogenesis in Vertebrate and Invertebrate Hosts.International journal of molecular sciences · 2020Review
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Reverse genetic systems are essential for the study of RNA viruses. Infectious clones remain the most widely used systems to manipulate viral genomes. Recently, a new PCR-based method called ISA (infectious subgenomic amplicons) has been developed. This approach has resulted in greater genetic diversity of the viral populations than that observed using infectious clone technology. However, for some studies, generation of clonal viral populations is necessary. In this study, we used the tick-borne encephalitis virus as model to demonstrate that utilization of a very high-fidelity, DNA-dependent DNA polymerase during the PCR step of the ISA procedure gives the possibility to reduce the genetic diversity of viral populations. We also concluded that the fidelity of the polymerase is not the only factor influencing this diversity. Studying the impact of genotype modification on virus phenotype is a crucial step for the development of reverse genetic methods. Here, we also demonstrated that the utilization of different PCR polymerases did not affect the phenotype (replicative fitness in cellulo and virulence in vivo) compared to the initial ISA procedure and the use of an infectious clone. In conclusion, we provide here an approach to control the genetic diversity of RNA viruses without modifying their phenotype.
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