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Reverse Genetics to Engineer Positive-Sense RNA Virus Variants
Published on: June 9, 2022
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A versatile reverse genetics platform for SARS-CoV-2 and other positive-strand RNA viruses
Alberto A Amarilla1, Julian D J Sng1, Rhys Parry1
1School of Chemistry and Molecular Biosciences, University of Queensland, St Lucia, QLD, Australia.
Nature Communications
|June 9, 2021
Summary
Researchers developed a novel method to synthesize infectious severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from its genetic material. This technique simplifies the creation of viable viruses for further study.
Area of Science:
- Virology
- Molecular Biology
- Infectious Diseases
Background:
- The COVID-19 pandemic is caused by SARS-CoV-2.
- Efficient methods for generating infectious viral particles are crucial for research and development.
Purpose of the Study:
- To develop a streamlined method for assembling full-length infectious viral cDNA.
- To demonstrate the utility of this method for SARS-CoV-2 and other viruses.
Main Methods:
- Circular polymerase extension reaction (CPER) methodology was employed.
- Overlapping cDNA fragments of viral RNA were assembled into a circular full-length cDNA with a promoter.
- Transfection of the circular cDNA into mammalian cells enabled virus recovery.
Main Results:
- Infectious SARS-CoV-2 was recovered from assembled circular cDNA.
- Recovered SARS-CoV-2 exhibited comparable in vitro and in vivo properties to the parental virus.
- CPER was successfully applied to generate other viruses, including Casuarina virus, Ross River virus, and Norovirus.
Conclusions:
- CPER is an efficient and versatile method for generating infectious full-length viral cDNA.
- This methodology facilitates the study of viral replication, virus-receptor interactions, and the development of reporter and mutant viruses.
