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Here, we report the use of synthetic genomics methodology to develop a reverse genetics system for ASFV using a CRISPR-Cas9\u2013inhibited 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.<\/jats:p>","DOI":"10.1126\/sciadv.adu7670","type":"journal-article","created":{"date-parts":[[2025,3,26]],"date-time":"2025-03-26T18:45:32Z","timestamp":1743014732000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark","source":"Crossref","is-referenced-by-count":8,"title":["A synthetic genomics-based African swine fever virus engineering platform"],"prefix":"10.1126","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7670-451X","authenticated-orcid":true,"given":"Walter","family":"Fuchs","sequence":"first","affiliation":[{"name":"Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5270-2272","authenticated-orcid":true,"given":"Nacyra","family":"Assad-Garcia","sequence":"additional","affiliation":[{"name":"J. 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