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FIPVs are suggested to emerge from feline enteric coronaviruses (FECVs) by acquiring mutations in specific genes in the course of persistent infections. Although numerous studies identified mutations predicted to be responsible for the FECV-FIPV biotype switch, the presumed roles of specific genetic changes in FIP pathogenesis have not been confirmed experimentally. Reverse genetics systems established previously for serotype I and the less common serotype II FCoVs were based on cell culture-adapted FIPV strains which, however, were shown to be unsuitable for FIP pathogenesis studies<jats:italic>in vivo<\/jats:italic>. To date, systems to produce and manipulate recombinant serotype I field viruses have not been developed, mainly because these viruses cannot be grown<jats:italic>in vitro<\/jats:italic>. Here, we report the first reverse genetics system based on a serotype I FECV field isolate that is suitable to produce high-titer stocks of recombinant FECVs. We demonstrate that these recombinant viruses cause productive persistent infections in cats that are similar to what is observed in natural infections. The system provides an excellent tool for studying FCoVs that do not grow in standard cell culture systems and will greatly facilitate studies into the molecular pathogenesis of FIP. Importantly, the system could also be adapted for studies of other RNA viruses with large genomes whose production and characterization<jats:italic>in vivo<\/jats:italic>are currently hampered by the lack of<jats:italic>in vitro<\/jats:italic>propagation systems.<\/jats:p><jats:p><jats:bold>IMPORTANCE<\/jats:bold>The availability of recombinant serotype I FCoV field isolates that are amenable to genetic manipulation is key to studying the molecular pathogenesis of FIP, especially since previous studies using cell culture-adapted FIPVs had proven unsuccessful. To our knowledge, we report the first serotype I FECV field isolate-based reverse genetics system that allows the production of high-titer recombinant virus stocks that can be used for subsequent<jats:italic>in vivo<\/jats:italic>studies in cats. The system represents a milestone in FCoV research. It provides an essential tool for studying the molecular pathogenesis of FIP and, more specifically, the functions of specific gene products in causing a fundamentally different progression of disease following acquisition of specific mutations. The system developed in this study will also be useful for studying other coronaviruses or more distantly related RNA viruses with large genomes for which suitable<jats:italic>in vitro<\/jats:italic>culture systems are not available.<\/jats:p>","DOI":"10.1128\/mbio.01422-18","type":"journal-article","created":{"date-parts":[[2018,7,30]],"date-time":"2018-07-30T09:59:22Z","timestamp":1532944762000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":28,"title":["Reverse Genetics for Type I Feline Coronavirus Field Isolate To Study the Molecular Pathogenesis of Feline Infectious Peritonitis"],"prefix":"10.1128","volume":"9","author":[{"given":"Rosina","family":"Ehmann","sequence":"first","affiliation":[{"name":"Institute of Virology, Justus Liebig University Giessen, Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Kristen-Burmann","sequence":"additional","affiliation":[{"name":"Institute of Virology, Justus Liebig University Giessen, Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Bank-Wolf","sequence":"additional","affiliation":[{"name":"Institute of Virology, Justus Liebig University Giessen, Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthias","family":"K\u00f6nig","sequence":"additional","affiliation":[{"name":"Institute of Virology, Justus Liebig University Giessen, Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christiane","family":"Herden","sequence":"additional","affiliation":[{"name":"Institute of Veterinary Pathology, Justus Liebig University Giessen, Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Torsten","family":"Hain","sequence":"additional","affiliation":[{"name":"Institute of Medical Microbiology, Justus Liebig University Giessen, Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heinz-J\u00fcrgen","family":"Thiel","sequence":"additional","affiliation":[{"name":"Institute of Virology, Justus Liebig University Giessen, Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Ziebuhr","sequence":"additional","affiliation":[{"name":"Institute of Medical Virology, Justus Liebig University Giessen, Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gergely","family":"Tekes","sequence":"additional","affiliation":[{"name":"Institute of Virology, Justus Liebig University Giessen, Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"785","volume-title":"Virus taxonomy: ninth report of the International Committee on Taxonomy of Viruses","author":"de Groot R","year":"2012","unstructured":"de Groot R, Cowley J, Enjuanes L, Faaberg K, Perlman S, Rottier P, Snijder E, Ziebuhr J, Gorbalenya A. 2012. 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