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ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Virology
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1572706
Design and Construction of a Fast Synthetic Modified Vaccinia Virus Ankara Reverse Genetics System for Advancing Vaccine Development
Provisionally accepted
Zhiqiang Gao
Busen Wang*
Tianyu Liu
Zhenghao Zhao
Jinghan Xu
Xiaofan Zhao
Zhe Zhang
Zuyuan Jia
Shipo WuYilong Yang
Wei Chen
Lihua Hou*- Beijing Institute of Biotechnology, Beijing, Beijing, China
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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.
Keywords: Modified Vaccinia virus Ankara (MVA)1, Reverse genetic system2, Rescue3, Viral vector Vaccine4, Viral genomes5
Received: 07 Feb 2025; Accepted: 07 Apr 2025.
Copyright: © 2025 Gao, Wang, Liu, Zhao, Xu, Zhao, Zhang, Jia, Wu, Yang, Chen and Hou. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Busen Wang, Beijing Institute of Biotechnology, Beijing, Beijing, China
Lihua Hou, Beijing Institute of Biotechnology, Beijing, Beijing, China
Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.