ArticleVirology journal2023
An optimized circular polymerase extension reaction-based method for functional analysis of SARS-CoV-2.
Article in Virology journal, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- SARS-CoV-2 infects olfactory neurons and basal stem cells and induces axonal degeneration through TRPV1 activation.iScience · 2026Article
- Cross-resistance patterns in SARS-CoV-2 against 3CL protease inhibitors.Nature communications · 2026Article
- Application of the CPER reverse genetics system for genetic engineering of rabies virus.Journal of virology · 2026Article
- Inhibition of coronaviral exoribonuclease activity by TRIM-mediated SUMOylation.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- Reverse genetics approach for arteriviruses using circular polymerase extension reaction.Access microbiology · 2026Article
- Assembly and Mutagenesis of Human Coronavirus OC43 Genomes in Yeast via Transformation-Associated Recombination.Bio-protocol · 2025Article
- Reverse genetics rescue of sylvatic dengue viruses.Journal of virology · 2025Article
- Two mutations in NS2B are responsible for attenuation of the yellow fever virus (YFV) vaccine strain 17D.PLoS pathogens · 2025Article
- Article
- Coronavirus endoribonuclease antagonizes ZBP1-mediated necroptosis and delays multiple cell death pathways.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- A yeast-based reverse genetics system to generate HCoV-OC43 reporter viruses encoding an eighth subgenomic RNA.Journal of virology · 2025Article
- Efficiency of reverse genetics methods for rescuing severe acute respiratory syndrome coronavirus 2.Journal of microbiology (Seoul, Korea) · 2025Article
- ISGylation of the SARS-CoV-2 N protein by HERC5 impedes N oligomerization and thereby viral RNA synthesis.Journal of virology · 2024Article
- ISGylation of the SARS-CoV-2 N protein by HERC5 impedes N oligomerization and thereby viral RNA synthesis.bioRxiv : the preprint server for biology · 2024Article
- Reverse genetics systems for SARS-CoV-2: Development and applications.Virologica Sinica · 2023Review
- Reverse Genetics Systems for Emerging and Re-Emerging Swine Coronaviruses and Applications.Viruses · 2023Review
- A viral pan-end RNA element and host complex define a SARS-CoV-2 regulon.Nature communications · 2023Article
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Abstract
backgroundReverse genetics systems have been crucial for studying specific viral genes and their relevance in the virus lifecycle, and become important tools for the rational attenuation of viruses and thereby for vaccine design. Recent rapid progress has been made in the establishment of reverse genetics systems for functional analysis of SARS-CoV-2, a coronavirus that causes the ongoing COVID-19 pandemic that has resulted in detrimental public health and economic burden. Among the different reverse genetics approaches, circular polymerase extension reaction (CPER) has become one of the leading methodologies to generate recombinant SARS-CoV-2 infectious clones. Although CPER has greatly facilitated SARS-CoV-2 analysis, it still has certain intrinsic limitations that impede the efficiency and robustness of virus rescue.
resultsWe developed an optimized CPER methodology which, through the use of a modified linker plasmid and by performing DNA nick ligation and direct transfection of permissive cells, overcomes certain intrinsic limitations of the 'traditional' CPER approaches for SARS-CoV-2, allowing for efficient virus rescue.
conclusionsThe herein described optimized CPER system may facilitate research studies to assess the contribution of SARS-CoV-2 genes and individual motifs or residues to virus replication, pathogenesis and immune escape, and may also be adapted to other viruses.
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