ArticleInfluenza and other respiratory viruses2023
The accuracy of reverse genetics systems for SARS-CoV-2: Circular polymerase extension reaction versus bacterial artificial chromosome.
Article in Influenza and other respiratory viruses, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- Application of the CPER reverse genetics system for genetic engineering of rabies virus.Journal of virology · 2026Article
- Article
- Assembly and Mutagenesis of Human Coronavirus OC43 Genomes in Yeast via Transformation-Associated Recombination.Bio-protocol · 2025Article
- Advances in the development of infectious clones of human coronaviruses and related applications.Biosafety and health · 2025Review
- Structural and virologic mechanism of the emergence of resistance to MProceedings of the National Academy of Sciences of the United States of America · 2024Article
- Reverse genetics systems for SARS-CoV-2: Development and applications.Virologica Sinica · 2023Review
- In vitro and in vivo characterization of SARS-CoV-2 resistance to ensitrelvir.Nature communications · 2023Article
- In vitro and in vivo characterization of SARS-CoV-2 strains resistant to nirmatrelvir.Nature communications · 2023Article
- The accuracy of reverse genetics systems for SARS-CoV-2: Circular polymerase extension reaction versus bacterial artificial chromosome.Influenza and other respiratory viruses · 2023Article
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3 authors.
Funding
Abstract
Background: Reverse genetics systems to rescue viruses from modified DNA are useful tools to investigate the molecular mechanisms of viruses. The COVID-19 pandemic prompted the development of several reverse genetics systems for SARS-CoV-2. The circular polymerase extension reaction (CPER) method enables the rapid generation of recombinant SARS-CoV-2; however, such PCR-based approaches could introduce unwanted mutations due to PCR errors. Methods: To compare the accuracy of CPER and a classic reverse genetics method using bacterial artificial chromosome (BAC), SARS-CoV-2 Wuhan/Hu-1/2019 was generated five times using BAC and five times using CPER. These 10 independent virus stocks were then deep sequencing, and the number of substitutions for which the frequency was greater than 10% was counted. Results: No nucleotide substitutions with a frequency of greater than 10% were observed in all five independent virus stocks generated by the BAC method. In contrast, three to five unwanted nucleotide substitutions with a frequency of more than 10% were detected in four of the five virus stocks generated by the CPER. Furthermore, four substitutions with frequencies greater than 20% were generated in three virus stocks by using the CPER. Conclusions: We found that the accuracy of the CPER method is lower than that of the BAC method. Our findings suggest care should be used when employing the CPER method.
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