Bibliographic Details
| Title: |
Monitoring of hepatitis E virus infection and replication by functional tagging of the ORF2 protein |
| Authors: |
Maliki Ankavay, Nathalie Da Silva, Angela Pollán, Noémie Oechslin, Katja Dinkelborg, Patrick Behrendt, Darius Moradpour, Jérôme Gouttenoire |
| Source: |
JHEP Reports, Vol 7, Iss 3, Pp 101293- (2025) |
| Publisher Information: |
Elsevier, 2025. |
| Publication Year: |
2025 |
| Collection: |
LCC:Diseases of the digestive system. Gastroenterology |
| Subject Terms: |
HEV, HiBiT, ORF2 protein, random insertion, split-luciferase, transposon, Diseases of the digestive system. Gastroenterology, RC799-869 |
| More Details: |
Background and Aims: Hepatitis E virus (HEV) infection is a leading cause of acute hepatitis worldwide. Understanding of the mechanisms underlying productive HEV infection remains incomplete and would benefit from technological advances improving current model systems. Methods: We exploited transposon-mediated random insertion and selection of viable clones to identify sites in the HEV open reading frame 2 (ORF2) protein, corresponding to the viral capsid, allowing for the insertion of reporter sequences in a functional context. Results: Short sequence insertions (5 amino acids) were tolerated at four distinct sites in the C-terminal region of the ORF2 protein, without significantly affecting viral capsid expression and subcellular localization as well as virus production. Full-length HEV genomes harboring larger sequence insertions such as an HA epitope tag, a highly sensitive miniaturized luciferase reporter (HiBiT) or a split GFP at these sites conserved their ability to produce infectious virus, with about a 1-log decrease in viral titers. Findings were confirmed in two different HEV genotype 3 clones. In addition, we demonstrate that HiBiT-tagged HEV, offering rapid and several-log amplitude detection, can be used for the evaluation of antiviral drugs and neutralizing antibodies. Conclusions: We describe a convenient, quantitative and potentially scalable system for the monitoring of HEV infection and replication in tissue culture. Impact and implications:: Hepatitis E virus infection is one of the most frequent causes of acute hepatitis and jaundice worldwide. As treatment options are limited and a vaccine is not universally available, the development of molecular tools to facilitate the identification of new therapeutic strategies is crucial. Based on a screening approach to identify viable insertion sites in the viral genome, we describe a versatile system for preparing recombinant viruses harboring split-reporter tags, i.e. luciferase and GFP. Proof-of-concept experiments revealed that convenient and quantitative monitoring of viral infection and replication is possible with this system, allowing for the evaluation of antiviral drugs and neutralizing antibodies. |
| Document Type: |
article |
| File Description: |
electronic resource |
| Language: |
English |
| ISSN: |
2589-5559 |
| Relation: |
http://www.sciencedirect.com/science/article/pii/S2589555924002970; https://doaj.org/toc/2589-5559 |
| DOI: |
10.1016/j.jhepr.2024.101293 |
| Access URL: |
https://doaj.org/article/f99ca0b6aeba4df1ba220481dc47a93a |
| Accession Number: |
edsdoj.f99ca0b6aeba4df1ba220481dc47a93a |
| Database: |
Directory of Open Access Journals |
