Design and construction of a fast synthetic modified vaccinia virus Ankara reverse genetics system for advancing vaccine development

The modified vaccinia virus Ankara (MVA) is approved for use as a smallpox and monkeypox virus vaccine and was also designed as a popular recombinant viral vector for vaccine development and gene therapy. However, the extensive genomes of poxviruses present a significant challenge for the developmen...

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Main Authors: Zhiqiang Gao, Busen Wang, Tianyu Liu, Zhenghao Zhao, Jinghan Xu, Xiaofan Zhao, Zhe Zhang, Zuyuan Jia, Yilong Yang, Shipo Wu, Wei Chen, Lihua Hou
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-04-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1572706/full
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author Zhiqiang Gao
Busen Wang
Tianyu Liu
Zhenghao Zhao
Jinghan Xu
Xiaofan Zhao
Zhe Zhang
Zuyuan Jia
Yilong Yang
Shipo Wu
Wei Chen
Lihua Hou
author_facet Zhiqiang Gao
Busen Wang
Tianyu Liu
Zhenghao Zhao
Jinghan Xu
Xiaofan Zhao
Zhe Zhang
Zuyuan Jia
Yilong Yang
Shipo Wu
Wei Chen
Lihua Hou
author_sort Zhiqiang Gao
collection DOAJ
description The modified vaccinia virus Ankara (MVA) is approved for use as a smallpox and monkeypox virus vaccine and was also designed as a popular recombinant viral vector for vaccine development and gene therapy. However, the extensive genomes of poxviruses present a significant challenge for the development of recombinant viral vaccines; therefore, it is essential to establish a user-friendly in vitro reverse genetic system. We systematically assembled the 180-kb MVA genome into a five-plasmid system, facilitating one-step packaging of the MVA virus. The MVA rescued using this system exhibited similar virological characteristics, including host cell tropism, growth kinetics, plaque size, and viral particles, comparable to those of wild-type MVA. Immunization with rescued MVA intramuscularly or subcutaneously triggered robust-specific immune responses and conferred protection against lethal attacks by the ectromelia virus in mice. We also developed a recombinant MVA-Luc-eGFP virus, which served as a tool for screening antiviral compounds against poxviruses. The synthetic MVA system efficiently generates recombinant vaccines with robust immune responses. These findings provide a novel and fast method for engineering large viral genomes with more specialized structures and lay a foundation for the advancement of more rapid and effective viral vector vaccines.
format Article
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institution OA Journals
issn 1664-302X
language English
publishDate 2025-04-01
publisher Frontiers Media S.A.
record_format Article
series Frontiers in Microbiology
spelling doaj-art-b175903a26ba4a0f855f6da334ea62ca2025-08-20T02:28:11ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-04-011610.3389/fmicb.2025.15727061572706Design and construction of a fast synthetic modified vaccinia virus Ankara reverse genetics system for advancing vaccine developmentZhiqiang GaoBusen WangTianyu LiuZhenghao ZhaoJinghan XuXiaofan ZhaoZhe ZhangZuyuan JiaYilong YangShipo WuWei ChenLihua HouThe modified vaccinia virus Ankara (MVA) is approved for use as a smallpox and monkeypox virus vaccine and was also designed as a popular recombinant viral vector for vaccine development and gene therapy. However, the extensive genomes of poxviruses present a significant challenge for the development of recombinant viral vaccines; therefore, it is essential to establish a user-friendly in vitro reverse genetic system. We systematically assembled the 180-kb MVA genome into a five-plasmid system, facilitating one-step packaging of the MVA virus. The MVA rescued using this system exhibited similar virological characteristics, including host cell tropism, growth kinetics, plaque size, and viral particles, comparable to those of wild-type MVA. Immunization with rescued MVA intramuscularly or subcutaneously triggered robust-specific immune responses and conferred protection against lethal attacks by the ectromelia virus in mice. We also developed a recombinant MVA-Luc-eGFP virus, which served as a tool for screening antiviral compounds against poxviruses. The synthetic MVA system efficiently generates recombinant vaccines with robust immune responses. These findings provide a novel and fast method for engineering large viral genomes with more specialized structures and lay a foundation for the advancement of more rapid and effective viral vector vaccines.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1572706/fullmodified vaccinia virus Ankara (MVA)reverse genetic systemrescueviral vector vaccineviral genomes
spellingShingle Zhiqiang Gao
Busen Wang
Tianyu Liu
Zhenghao Zhao
Jinghan Xu
Xiaofan Zhao
Zhe Zhang
Zuyuan Jia
Yilong Yang
Shipo Wu
Wei Chen
Lihua Hou
Design and construction of a fast synthetic modified vaccinia virus Ankara reverse genetics system for advancing vaccine development
Frontiers in Microbiology
modified vaccinia virus Ankara (MVA)
reverse genetic system
rescue
viral vector vaccine
viral genomes
title Design and construction of a fast synthetic modified vaccinia virus Ankara reverse genetics system for advancing vaccine development
title_full Design and construction of a fast synthetic modified vaccinia virus Ankara reverse genetics system for advancing vaccine development
title_fullStr Design and construction of a fast synthetic modified vaccinia virus Ankara reverse genetics system for advancing vaccine development
title_full_unstemmed Design and construction of a fast synthetic modified vaccinia virus Ankara reverse genetics system for advancing vaccine development
title_short Design and construction of a fast synthetic modified vaccinia virus Ankara reverse genetics system for advancing vaccine development
title_sort design and construction of a fast synthetic modified vaccinia virus ankara reverse genetics system for advancing vaccine development
topic modified vaccinia virus Ankara (MVA)
reverse genetic system
rescue
viral vector vaccine
viral genomes
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1572706/full
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