{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T04:58:42Z","timestamp":1790830722700,"version":"4.1.0"},"reference-count":60,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T00:00:00Z","timestamp":1790553600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004704","name":"National Research Council of Thailand","doi-asserted-by":"publisher","award":["N42A670624"],"award-info":[{"award-number":["N42A670624"]}],"id":[{"id":"10.13039\/501100004704","id-type":"DOI","asserted-by":"publisher"}]},{"award":["N42A670624"],"award-info":[{"award-number":["N42A670624"]}],"id":[{"id":"https:\/\/ror.org\/018wfhg78","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004539","name":"Kasetsart University","doi-asserted-by":"publisher","award":["FF(KU)16.70"],"award-info":[{"award-number":["FF(KU)16.70"]}],"id":[{"id":"10.13039\/501100004539","id-type":"DOI","asserted-by":"publisher"}]},{"award":["FF(KU)16.70"],"award-info":[{"award-number":["FF(KU)16.70"]}],"id":[{"id":"https:\/\/ror.org\/05gzceg21","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Feline infectious peritonitis virus (FIPV) causes a fatal disease in cats that remains challenging to treat due to the absence of effective vaccines and limited antiviral therapies. Given its central role in viral replication and transcription, RNA-dependent RNA polymerase (RdRp) represents a highly promising and strategic target for antiviral drug development. To enable live-cell monitoring of RdRp activity without external substrates or cell disruption, we developed a biologically validated, cell-based dual-fluorescent reporter assay using green fluorescent protein and red fluorescent protein to evaluate RdRp-dependent reporter activity. Following validation with the well-characterized RdRp inhibitor ribavirin, the dual-reporter platform achieved a Z-factor of 0.54 and a Z\u2032-factor of 0.61. The system was further utilized to assess the antiviral potential of fifteen alkaloid compounds. Among compounds evaluated, lycorine and lycorine hydrochloride exhibited potent inhibition of RdRp-dependent reporter activity and yielded IC50 values of 2.83 \u00b1 0.45 \u03bcM and 1.58 \u00b1 0.20 \u03bcM, respectively. Further evaluation revealed that both compounds significantly reduced FIPV replication in infected cell. These findings identify lycorine and lycorine hydrochloride as promising anti-FIPV candidates and support further investigation of lycorine-class alkaloids as anti-coronaviral scaffolds. Together, the validated cell-based screening platform and whole-virus antiviral activity provide a foundation for advancing RdRp-targeted antiviral discovery against FIPV.<\/jats:p>","DOI":"10.3390\/ijms27198682","type":"journal-article","created":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T16:50:54Z","timestamp":1790614254000},"page":"8682","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and Application of a Dual-Fluorescent Reporter Assay for Feline Infectious Peritonitis Virus RNA-Dependent RNA Polymerase Inhibitor Screening"],"prefix":"10.3390","volume":"27","author":[{"given":"Ploypailin","family":"Semkum","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-5953-976X","authenticated-orcid":false,"given":"Varanya","family":"Lueangaramkul","sequence":"additional","affiliation":[{"name":"Department of Anatomy, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Supichaya","family":"Jiwattanasap","sequence":"additional","affiliation":[{"name":"Graduate Program in Animal Health and Biomedical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-8162-8563","authenticated-orcid":false,"given":"Natjira","family":"Mana","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-6542-6667","authenticated-orcid":false,"given":"Nantawan","family":"Phetcharat","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5595-3296","authenticated-orcid":false,"given":"Porntippa","family":"Lekcharoensuk","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6912-9764","authenticated-orcid":false,"given":"Sirin","family":"Theerawatanasirikul","sequence":"additional","affiliation":[{"name":"Department of Anatomy, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"199059","DOI":"10.1016\/j.virusres.2023.199059","article-title":"An updated review of feline coronavirus: Mind the two biotypes","volume":"326","author":"Gao","year":"2023","journal-title":"Virus Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.tvjl.2014.04.017","article-title":"An update on feline infectious peritonitis: Virology and immunopathogenesis","volume":"201","author":"Pedersen","year":"2014","journal-title":"Vet. 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