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A reverse genetics system of African horse sickness virus reveals existence of primary replication.
Abstract: African horse sickness virus (AHSV), a member of the orbivirus genus of the family Reoviridae, is an insect-vectored pathogen of horses of concern to the equine industry. Studies on AHSV replication and pathogenesis have been hampered by the lack of reverse genetics allowing targeted mutation of viral genomes. We demonstrate that AHSV single-stranded RNA synthesized in vitro (core transcripts) is infectious and that there are distinct primary and secondary stages of the replication cycle. Transfection with a mixture of core transcripts from two different serotypes or a mixture of core transcripts and a T7 derived transcript resulted in the recovery of reassortant viruses. Recovery of infectious ASHV from nucleic acid will benefit investigation of the virus and the generation of attenuated vaccines.
Copyright (c) 2010 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
Publication Date: 2010-06-26 PubMed ID: 20600010DOI: 10.1016/j.febslet.2010.06.030Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The researchers’ work focuses on the African horse sickness virus (AHSV), a deadly virus affecting horses. They developed a reverse genetics system that allows them to understand the replication and pathogenesis of AHSV by generating targeted mutations on its genomes.
Understanding AHSV and Its Significance
- The study explores the African horse sickness virus (AHSV), which belongs to the orbivirus genus of the family Reoviridae. This virus is a significant concern to the equine industry as it is a pathogen of horses transmitted by insects.
- Understanding this virus’ characteristics, replication process, and pathogenesis is vital due to its impact on the equine industry. However, these studies have been obstructed due to the absence of reverse genetics, a method needed to generate targeted viral genome mutations.
Introduction of a Reverse Genetics System
- The scientists developed a reverse genetics system for AHSV. Reverse genetics is an innovative process that permits scientists to comprehend the function of a gene by analyzing the phenotypic effects of specific engineered gene sequences. Using this method, they could explore the impact of targeted alterations or mutations in the AHSV genomes.
- They further exhibited that the AHSV single-stranded RNA synthesized in vitro (core transcripts) is infectious. This suggests that AHSV can be propagated in an artificial environment outside of its natural host, an essential step in understanding the virus better.
Stages of AHSV Replication Cycle
- The research identified primary and secondary stages of the AHSV replication cycle. This could provide critical insights into how the virus proliferates and the stages at which it could potentially be interrupted in future research.
- Through transfection, they mixed core transcripts from two different serotypes or a blend of core transcripts and a T7 derived transcript that resulted in the recovery of reassortant viruses. This demonstrates the mechanism by which AHSV possibly evolves and adapts through recombination events.
Future Applications of the Research
- The successful recovery of infectious AHSV from nucleic acid will open new avenues for further exploration of the virus’s nature and behaviors. This is a crucial step for the development of strategies against AHSV.
- Moreover, this research could pave the way for the production of attenuated vaccines. An attenuated vaccine utilizes a weakened form of the germ that causes a disease; hence, understanding the genetic manipulation of AHSV may lead to the creation of a more effective vaccine against it.
Cite This Article
APA
Matsuo E, Celma CC, Roy P.
(2010).
A reverse genetics system of African horse sickness virus reveals existence of primary replication.
FEBS Lett, 584(15), 3386-3391.
https://doi.org/10.1016/j.febslet.2010.06.030 Publication
Researcher Affiliations
- Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, UK.
MeSH Terms
- African Horse Sickness Virus / genetics
- African Horse Sickness Virus / isolation & purification
- African Horse Sickness Virus / physiology
- African Horse Sickness Virus / ultrastructure
- Animals
- Cell Line
- DNA, Complementary / genetics
- Gene Expression Regulation, Viral
- Genetic Techniques
- Genome, Viral / genetics
- RNA, Messenger / genetics
- RNA, Messenger / metabolism
- RNA, Viral / genetics
- Reassortant Viruses / classification
- Reassortant Viruses / genetics
- Reassortant Viruses / physiology
- Serotyping
- Transfection
- Virion / isolation & purification
- Virion / ultrastructure
- Virus Replication / physiology
Grant Funding
- BB/F007159/1 / Biotechnology and Biological Sciences Research Council
- BB/F02049X/1 / Biotechnology and Biological Sciences Research Council
Citations
This article has been cited 18 times.- Pitchers KG, Boakye OD, Campeotto I, Daly JM. The Potential of Plant-Produced Virus-like Particle Vaccines for African Horse Sickness and Other Equine Orbiviruses. Pathogens 2024 May 28;13(6).
- Calvo-Pinilla E, Marín-López A, Utrilla-Trigo S, Jiménez-Cabello L, Ortego J. Reverse genetics approaches: a novel strategy for African horse sickness virus vaccine design. Curr Opin Virol 2020 Oct;44:49-56.
- Dennis SJ, Meyers AE, Hitzeroth II, Rybicki EP. African Horse Sickness: A Review of Current Understanding and Vaccine Development. Viruses 2019 Sep 11;11(9).
- Desselberger U. The 13th International Double-Stranded RNA Virus Symposium, Houffalize, Belgium, 24 to 28 September 2018. J Virol 2019 Feb 15;93(4).
- Schade-Weskott ML, van Schalkwyk A, Koekemoer JJO. A correlation between capsid protein VP2 and the plaque morphology of African horse sickness virus in cell culture. Virus Genes 2018 Aug;54(4):527-535.
- Mathebula EM, Faber FE, Van Wyngaardt W, Van Schalkwyk A, Pretorius A, Fehrsen J. B-cell epitopes of African horse sickness virus serotype 4 recognised by immune horse sera. Onderstepoort J Vet Res 2017 Feb 24;84(1):e1-e12.
- Fajardo T Jr, AlShaikhahmed K, Roy P. Generation of infectious RNA complexes in Orbiviruses: RNA-RNA interactions of genomic segments. Oncotarget 2016 Nov 8;7(45):72559-72570.
- Lulla V, Lulla A, Wernike K, Aebischer A, Beer M, Roy P. Assembly of Replication-Incompetent African Horse Sickness Virus Particles: Rational Design of Vaccines for All Serotypes. J Virol 2016 Aug 15;90(16):7405-7414.
- Matsuo E, Saeki K, Roy P, Kawano J. Development of reverse genetics for Ibaraki virus to produce viable VP6-tagged IBAV. FEBS Open Bio 2015;5:445-53.
- Burkhardt C, Sung PY, Celma CC, Roy P. Structural constraints in the packaging of bluetongue virus genomic segments. J Gen Virol 2014 Oct;95(Pt 10):2240-2250.
- Patel A, Roy P. The molecular biology of Bluetongue virus replication. Virus Res 2014 Mar;182:5-20.
- Richards JE, Desselberger U, Lever AM. Experimental pathways towards developing a rotavirus reverse genetics system: synthetic full length rotavirus ssRNAs are neither infectious nor translated in permissive cells. PLoS One 2013;8(9):e74328.
- Matsuo E, Roy P. Minimum requirements for bluetongue virus primary replication in vivo. J Virol 2013 Jan;87(2):882-9.
- Trask SD, Boehme KW, Dermody TS, Patton JT. Comparative analysis of Reoviridae reverse genetics methods. Methods 2013 Feb;59(2):199-206.
- Tanaka T, Eusebio-Cope A, Sun L, Suzuki N. Mycoreovirus genome alterations: similarities to and differences from rearrangements reported for other reoviruses. Front Microbiol 2012;3:186.
- Manole V, Laurinmäki P, Van Wyngaardt W, Potgieter CA, Wright IM, Venter GJ, van Dijk AA, Sewell BT, Butcher SJ. Structural insight into African horsesickness virus infection. J Virol 2012 Aug;86(15):7858-66.
- Matsuo E, Roy P. Bluetongue virus VP1 polymerase activity in vitro: template dependency, dinucleotide priming and cap dependency. PLoS One 2011;6(11):e27702.
- Trask SD, Taraporewala ZF, Boehme KW, Dermody TS, Patton JT. Dual selection mechanisms drive efficient single-gene reverse genetics for rotavirus. Proc Natl Acad Sci U S A 2010 Oct 26;107(43):18652-7.
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