ArticleThe Journal of general virology2017
Transgene expression in the genome of Middle East respiratory syndrome coronavirus based on a novel reverse genetics system utilizing Red-mediated recombination cloning.
Article in The Journal of general virology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 18 citations in OpenAlex.
- MERS-CoV accessory ORF proteins in innate immune evasion: an evidence-graded functional framework.Frontiers in immunology · 2026Review
- Identification of an optimal exogenous gene insertion site (P-M) and establishment of a reverse genetics system for aMPV/A.PloS one · 2026Article
- Review
- Endogenous IFITMs boost SARS-coronavirus 1 and 2 replication whereas overexpression inhibits infection by relocalizing ACE2.iScience · 2023Article
- Human lungs show limited permissiveness for SARS-CoV-2 due to scarce ACE2 levels but virus-induced expansion of inflammatory macrophages.The European respiratory journal · 2022Article
- Reverse Genetics with a Full-Length Infectious cDNA Clone of Bovine Torovirus.Journal of virology · 2022Article
- Optimization of a Lambda-RED Recombination Method for Rapid Gene Deletion in Human Cytomegalovirus.International journal of molecular sciences · 2021Article
- Functional comparison of MERS-coronavirus lineages reveals increased replicative fitness of the recombinant lineage 5.Nature communications · 2021Article
- Recent Progress in Torovirus Molecular Biology.Viruses · 2021Review
- Article
- Toward Understanding Molecular Bases for Biological Diversification of Human Coronaviruses: Present Status and Future Perspectives.Frontiers in microbiology · 2020Review
- Reverse genetic systems: Rational design of coronavirus live attenuated vaccines with immune sequelae.Advances in virus research · 2020Review
- SKP2 attenuates autophagy through Beclin1-ubiquitination and its inhibition reduces MERS-Coronavirus infection.Nature communications · 2019Article
- Article
- Article
- Detection of distinct MERS-Coronavirus strains in dromedary camels from Kenya, 2017.Emerging microbes & infections · 2018Article
Corrections and comments
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Authors and funding
7 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Middle East respiratory syndrome coronavirus (MERS-CoV) is a high-priority pathogen in pandemic preparedness research. Reverse genetics systems are a valuable tool to study viral replication and pathogenesis, design attenuated vaccines and create defined viral assay systems for applications such as antiviral screening. Here we present a novel reverse genetics system for MERS-CoV that involves maintenance of the full-length viral genome as a cDNA copy inserted in a bacterial artificial chromosome amenable to manipulation by homologue recombination, based on the bacteriophage λ Red recombination system. Based on a full-length infectious MERS-CoV cDNA clone, optimal genomic insertion sites and expression strategies for GFP were identified and used to generate a reporter MERS-CoV expressing GFP in addition to the complete set of viral proteins. GFP was genetically fused to the N-terminal part of protein 4a, from which it is released during translation via porcine teschovirus 2A peptide activity. The resulting reporter virus achieved titres nearly identical to the wild-type virus 48 h after infection of Vero cells at m.o.i. 0.001 (1×10
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