Fuchs Walter, Assad-Garcia Nacyra, Abkallo Hussein M, Xue Yong, Oldfield Lauren M, Fedorova Nadia, Hübner Alexandra, Kabuuka Tonny, Pannhorst Katrin, Höper Dirk, Nene Vishvanath, Gonzalez-Juarbe Norberto, Steinaa Lucilla, Vashee Sanjay
Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, Germany.
J. Craig Venter Institute, Rockville, MD 20850, USA.
Sci Adv. 2025 Mar 28;11(13):eadu7670. doi: 10.1126/sciadv.adu7670. Epub 2025 Mar 26.
African swine fever (ASF) is a deadly viral disease in domestic pigs that has a large global economic impact for the swine industry. It is present in Africa, Europe, Asia, and in the Caribbean island of Hispaniola. There are no effective treatments or broadly licensed vaccines to prevent disease. Efforts to counteract ASF have been hampered because of the lack of convenient tools to engineer its etiological agent, ASF virus (ASFV), largely due to its large noninfectious genome. Here, we report the use of synthetic genomics methodology to develop a reverse genetics system for ASFV using a CRISPR-Cas9-inhibited self-helper virus to reconstitute live recombinant ASFV from synthetic genomes to rapidly generate a variety of combinatorial mutants of ASFV. The method will substantially facilitate the development of therapeutics or subunit and live-attenuated vaccines for ASF. This synthetic genomics-based approach has wide-ranging impact because it can be applied to rapidly develop reverse genetics tools for emerging viruses with noninfectious genomes.
非洲猪瘟（ASF）是家猪的一种致命病毒性疾病，对全球养猪业造成巨大经济影响。它在非洲、欧洲、亚洲以及加勒比海的伊斯帕尼奥拉岛存在。目前没有有效的治疗方法或广泛许可的疫苗来预防该疾病。由于缺乏便捷的工具来改造其病原体非洲猪瘟病毒（ASFV），对抗ASF的努力受到阻碍，这主要是因为其基因组庞大且无感染性。在此，我们报告使用合成基因组学方法开发了一种针对ASFV的反向遗传学系统，该系统利用一种受CRISPR-Cas9抑制的自我辅助病毒从合成基因组中重建活的重组ASFV，从而快速产生多种ASFV组合突变体。该方法将极大地促进针对ASF的治疗药物或亚单位及减毒活疫苗的开发。这种基于合成基因组学的方法具有广泛影响，因为它可应用于快速开发针对具有无感染性基因组的新兴病毒的反向遗传学工具。
