{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:11:32Z","timestamp":1760145092858,"version":"build-2065373602"},"reference-count":31,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2024,6,19]],"date-time":"2024-06-19T00:00:00Z","timestamp":1718755200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Research Foundation of Korea","award":["2022R1I1A2066092","2022R1C1C2004274","2020R1A5A2017323","5199990614732"],"award-info":[{"award-number":["2022R1I1A2066092","2022R1C1C2004274","2020R1A5A2017323","5199990614732"]}]},{"name":"Korean ministry of education","award":["2022R1I1A2066092","2022R1C1C2004274","2020R1A5A2017323","5199990614732"],"award-info":[{"award-number":["2022R1I1A2066092","2022R1C1C2004274","2020R1A5A2017323","5199990614732"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>MHV-A59 is a beta-coronavirus that causes demyelinating encephalitis and hepatitis in mice. Recently, the mouse infection model of MHV-A59 has been used as an alternative animal infection model for SARS-CoV and SARS-CoV-2, aiding the development of new antiviral drugs. In this study, the MHV-A59 model was employed to investigate the potential of SARS-CoV-2 UTRs as new targets for antiviral drugs. Optimal targets within the MHV-A59 UTRs were identified using a shRNA and siRNA design tool, focusing on RNA secondary stem-loop (SL) structures in the UTRs. We then examined whether the designed RNAi constructs could inhibit MHV-A59 replication. In the 5\u2032UTR, the stem-loop 1 (SL1) was identified as the most effective target, while in the 3\u2032UTR, the minimal element for the initiation of negative-strand RNA synthesis (MIN) proved to be the most effective. Importantly, siRNAs targeting SL1 and MIN structures significantly reduced total RNA synthesis, negative-strand genomic RNA synthesis, subgenomic (sg) RNA synthesis, viral titer, and the plaque size of MHV-A59 compared to the control. Although not statistically significant, the combination of siSL1 and siMIN had a stronger effect on inhibiting MHV-A59 replication than either siRNA monotherapy. Interestingly, while the SL1 structure is present in both MHV and SARS-CoV-2, the MIN structure is unique to MHV. Thus, the SL1 of SARS-CoV-2 may represent a novel and promising target for RNAi-based antiviral drugs.<\/jats:p>","DOI":"10.3390\/pathogens13060518","type":"journal-article","created":{"date-parts":[[2024,6,19]],"date-time":"2024-06-19T11:47:17Z","timestamp":1718797637000},"page":"518","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Evaluation of RNA Secondary Stem-Loop Structures in the UTRs of Mouse Hepatitis Virus as New Therapeutic Targets"],"prefix":"10.3390","volume":"13","author":[{"given":"Gyuhyun","family":"Kang","sequence":"first","affiliation":[{"name":"Vessel-Organ Interaction Research Center, Research Institute of Pharmaceutical Science, College of Pharmacy, Kyungpook National University, Daegu 41566, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sun Hee","family":"Lee","sequence":"additional","affiliation":[{"name":"Vessel-Organ Interaction Research Center, Research Institute of Pharmaceutical Science, College of Pharmacy, Kyungpook National University, Daegu 41566, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Miyeon","family":"Cho","sequence":"additional","affiliation":[{"name":"Vessel-Organ Interaction Research Center, Research Institute of Pharmaceutical Science, College of Pharmacy, Kyungpook National University, Daegu 41566, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ji-hyeon","family":"Kim","sequence":"additional","affiliation":[{"name":"Vessel-Organ Interaction Research Center, Research Institute of Pharmaceutical Science, College of Pharmacy, Kyungpook National University, Daegu 41566, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9131-5742","authenticated-orcid":false,"given":"Hyosun","family":"Cho","sequence":"additional","affiliation":[{"name":"Duksung Innovative Drug Center, College of Pharmacy, Duksung Women\u2019s University, Seoul 01369, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0622-5839","authenticated-orcid":false,"given":"Hyojeung","family":"Kang","sequence":"additional","affiliation":[{"name":"Vessel-Organ Interaction Research Center, Research Institute of Pharmaceutical Science, College of Pharmacy, Kyungpook National University, Daegu 41566, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,6,19]]},"reference":[{"key":"ref_1","first-page":"3","article-title":"Properties of coronavirus and SARS-CoV-2","volume":"42","author":"Malik","year":"2020","journal-title":"Malays. 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