{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T16:52:53Z","timestamp":1785430373949,"version":"3.56.0"},"reference-count":96,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,6,9]],"date-time":"2026-06-09T00:00:00Z","timestamp":1780963200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["82402625"],"award-info":[{"award-number":["82402625"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"award":["82402625"],"award-info":[{"award-number":["82402625"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Rotaviruses remain the leading cause of severe dehydrating diarrhea and associated mortality in children under five years of age worldwide. The successful global rollout of live-attenuated oral rotavirus vaccines has dramatically reduced rotavirus gastroenteritis morbidity and mortality, unequivocally demonstrating their excellent safety profile and potent induction of mucosal immunity. These attributes highlight the substantial potential of rotaviruses as novel oral mucosal vaccine vectors. Recent breakthroughs in reverse genetics, particularly the establishment of a fully plasmid-based system in 2017, have enabled precise insertion and stable expression of foreign antigens at targeted genomic loci. This advance has opened a viable pathway for developing multivalent oral mucosal vaccines. This review traces the historical development of rotavirus reverse genetics and summarizes the latest progress in its application as a vaccine vector platform. We focus on key strategies for foreign gene insertion and immunological outcomes in animal models, while critically evaluating persistent challenges\u2014virus rescue efficiency, genetic stability of inserts, and limitations of current animal models\u2014and outlining the rational design framework for RV-based vectors with improved stability, expression efficiency, and immunogenicity.<\/jats:p>","DOI":"10.3390\/v18060655","type":"journal-article","created":{"date-parts":[[2026,6,11]],"date-time":"2026-06-11T05:50:58Z","timestamp":1781157058000},"page":"655","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Reprogramming Rotavirus: Reverse Genetics-Driven Design of Viral Vector Platforms"],"prefix":"10.3390","volume":"18","author":[{"given":"Ke","family":"Li","sequence":"first","affiliation":[{"name":"NHC Key Laboratory of Medical Virology and Viral Disease, National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao","family":"Wei","sequence":"additional","affiliation":[{"name":"NHC Key Laboratory of Medical Virology and Viral Disease, National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"},{"name":"School of Public Health, Gansu University of Traditional Chinese Medicine, Lanzhou 730000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuanze","family":"Ouyang","sequence":"additional","affiliation":[{"name":"NHC Key Laboratory of Medical Virology and Viral Disease, National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiafei","family":"Liu","sequence":"additional","affiliation":[{"name":"The First Clinical Medical Institute, Henan University of Chinese Medicine, Zhengzhou 450000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-1995-6021","authenticated-orcid":false,"given":"Pengdi","family":"Chai","sequence":"additional","affiliation":[{"name":"NHC Key Laboratory of Medical Virology and Viral Disease, National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"},{"name":"CAS Key Laboratory of Pathogen Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Bai","sequence":"additional","affiliation":[{"name":"Center for Drug Evaluation, National Medical Products Administration, Beijing 100163, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3090-4150","authenticated-orcid":false,"given":"Zhaojun","family":"Duan","sequence":"additional","affiliation":[{"name":"NHC Key Laboratory of Medical Virology and Viral Disease, National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.virusres.2014.06.016","article-title":"Rotaviruses","volume":"190","author":"Desselberger","year":"2014","journal-title":"Virus Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"958","DOI":"10.1001\/jamapediatrics.2018.1960","article-title":"Rotavirus Vaccination and the Global Burden of Rotavirus Diarrhea Among Children Younger Than 5 Years","volume":"172","author":"Troeger","year":"2018","journal-title":"JAMA Pediatr."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/S1473-3099(19)30401-3","article-title":"Quantifying risks and interventions that have affected the burden of diarrhoea among children younger than 5 years: An analysis of the Global Burden of Disease Study 2017","volume":"20","author":"Troeger","year":"2020","journal-title":"Lancet Infect. 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