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Updated: Jan 15, 2026
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Reverse Genetics to Engineer Positive-Sense RNA Virus Variants
Published on: June 9, 2022
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First Versatile Reverse Genetics System for DNA Viruses Using Circular Polymerase Extension Reaction.
Hirotaka Yamamoto1, Tomokazu Tamura1,2,3,4,5, Rigel Suzuki1,2,4
1Department of Microbiology and Immunology, Faculty of Medicine, Hokkaido University, Sapporo, Japan.
Microbiology and Immunology
|October 13, 2025
Summary
This study introduces circular polymerase extension reaction (CPER) for rapid DNA virus reverse genetics. The method efficiently generates infectious recombinant adenoviruses (AdVs), accelerating AdV research and therapeutic development.
Area of Science:
- Virology
- Molecular Biology
- Genetic Engineering
Background:
- Reverse genetics is crucial for virus research but conventional methods for DNA viruses are complex.
- Circular polymerase extension reaction (CPER) is established for RNA viruses but not DNA viruses.
Purpose of the Study:
- To adapt and apply CPER for efficient generation of infectious recombinant DNA viruses.
- To evaluate CPER as a novel reverse genetics platform for adenoviruses (AdVs).
Main Methods:
- Applied CPER to generate full-length cDNA of two adenovirus serotypes.
- Assessed the infectivity and replication of generated recombinant AdVs.
- Confirmed viral replication using immunostaining techniques.
Main Results:
- Successfully generated infectious recombinant adenoviruses (AdVs) using CPER.
- Recombinant AdVs exhibited replication comparable to parental strains.
- CPER proved to be a rapid and efficient method for DNA virus reverse genetics.
Conclusions:
- CPER is a viable and efficient platform for reverse genetics of DNA viruses.
- This method can accelerate adenovirus research and the development of AdV-based therapeutics.
- The findings open new avenues for genetic manipulation of DNA viruses.
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