{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T18:17:57Z","timestamp":1784139477105,"version":"3.55.0"},"reference-count":26,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T00:00:00Z","timestamp":1782345600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The molecular mechanisms governing the efficient packaging of the large SARS-CoV-2 RNA genome into progeny virions remain incompletely understood, with the role of untranslated regions (UTRs) being particularly enigmatic. Leveraging proximity ligation sequencing data, we identified direct, high-frequency interactions between the viral packaging signal PS9 and both the 5\u2032 and 3\u2032 UTRs during intracellular replication stages. Functional validation using an infectious virus-like particle (iVLP) system demonstrated that genomes incorporating SARS-CoV-2 UTRs exhibited significantly enhanced packaging efficiency, yielding an increase in both packaged RNA copies and reporter gene expression post-infection. Competitive packaging assays confirmed the UTRs confer a selective advantage during particle assembly. Mechanistically, Western blot and digital Western analysis revealed that UTR-containing iVLPs incorporated approximately 2-fold more nucleocapsid (N) proteins, suggesting enhanced N recruitment or retention. The deletion of specific core sequences within the UTRs predicted to form a base pair with PS9 abrogated this enhancement, suggesting the functional significance of the UTR-PS9 interaction interface. Collectively, these results establish that the 5\u2032 and 3\u2032 UTRs act synergistically through direct RNA-RNA interactions with PS9 to promote N protein recruitment and enhance packaging efficiency in a PS9-dependent iVLPs system. This UTR-PS9 regulatory axis presents a novel target for therapeutic intervention against SARS-CoV-2 and related coronaviruses.<\/jats:p>","DOI":"10.3390\/v18070700","type":"journal-article","created":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T08:14:12Z","timestamp":1782375252000},"page":"700","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Involvement of 5\u2032 and 3\u2032 UTRs in SARS-CoV-2 Virus-like Particle Genome Packaging"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6088-2293","authenticated-orcid":false,"given":"Zhang","family":"Zhang","sequence":"first","affiliation":[{"name":"Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kun","family":"Yang","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Henan Normal University, Xinxiang 453007, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fangze","family":"Shao","sequence":"additional","affiliation":[{"name":"Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenlong","family":"Shen","sequence":"additional","affiliation":[{"name":"Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1872-1546","authenticated-orcid":false,"given":"Ping","family":"Li","sequence":"additional","affiliation":[{"name":"Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yue","family":"Zhang","sequence":"additional","affiliation":[{"name":"Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5456-837X","authenticated-orcid":false,"given":"Junjie","family":"Xu","sequence":"additional","affiliation":[{"name":"Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dejian","family":"Xie","sequence":"additional","affiliation":[{"name":"Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6502-3842","authenticated-orcid":false,"given":"Chudong","family":"Wang","sequence":"additional","affiliation":[{"name":"Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4124-1359","authenticated-orcid":false,"given":"Guoying","family":"Yu","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Henan Normal University, Xinxiang 453007, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9650-112X","authenticated-orcid":false,"given":"Jun","family":"Zhang","sequence":"additional","affiliation":[{"name":"Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhihu","family":"Zhao","sequence":"additional","affiliation":[{"name":"Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4006-663X","authenticated-orcid":false,"given":"Yan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.coviro.2021.11.011","article-title":"A Structural View of the SARS-CoV-2 Virus and Its Assembly","volume":"52","author":"Hardenbrook","year":"2022","journal-title":"Curr. 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