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Updated: Jul 4, 2025
13:04
A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
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Virus-derived circular RNAs populate hepatitis C virus-infected cells
Qian M Cao1, Pakpoom Boonchuen2,3, Tzu-Chun Chen1
1Department of Microbiology & Immunology, School of Medicine, Stanford University, Stanford, CA 94305.
Summary
Hepatitis C virus RNA can form circular RNAs (circRNAs) that enhance viral replication. These novel virus-derived circRNAs (vcircRNAs) can be translated or boost viral RNA levels, impacting cellular pathways.
Area of Science:
- Virology
- Molecular Biology
- RNA Biology
Background:
- Eukaryotic pre-mRNAs undergo backsplicing to form stable circular RNAs (circRNAs).
- circRNAs can sequester proteins or small RNAs, influencing cellular pathways.
- Viruses often co-opt host cell machinery for their replication.
Purpose of the Study:
- To investigate if the hepatitis C virus (HCV) RNA genome is processed into virus-derived circRNAs (vcircRNAs).
- To determine the functional roles of potential vcircRNAs in HCV infection.
Main Methods:
- Computational analysis of RNA-sequencing data to predict vcircRNA formation.
- Experimental verification using rolling-circle amplification.
- Assays to assess translation and effects on viral RNA abundance.
Main Results:
- Hundreds of vcircRNAs were computationally predicted from the HCV RNA genome.
- Over a dozen vcircRNAs were experimentally confirmed.
- vcircRNAs containing the internal ribosome entry site were translated into proviral proteins.
- Two abundant, non-translated vcircRNAs enhanced viral RNA levels.
Conclusions:
- HCV RNA can be processed into diverse vcircRNAs.
- vcircRNAs represent a novel mechanism modulating HCV amplification.
- These findings reveal new roles for circRNAs in RNA virus infections.
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