{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T13:45:46Z","timestamp":1782827146585,"version":"3.54.5"},"posted":{"date-parts":[[2026,6,27]]},"group-title":"Microbiology","reference-count":69,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,6,27]],"date-time":"2026-06-27T00:00:00Z","timestamp":1782518400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["R00-AI159049"],"award-info":[{"award-number":["R00-AI159049"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["R01-AI186810"],"award-info":[{"award-number":["R01-AI186810"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["R01-AI169537"],"award-info":[{"award-number":["R01-AI169537"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["R01-AI140442"],"award-info":[{"award-number":["R01-AI140442"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["T32-AI007632"],"award-info":[{"award-number":["T32-AI007632"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"name":"National Cancer Institute","award":["T32-CA009171"],"award-info":[{"award-number":["T32-CA009171"]}]},{"name":"National Cancer Institute","award":["P30-CA010815"],"award-info":[{"award-number":["P30-CA010815"]}]},{"DOI":"10.13039\/100001340","name":"W. W. Smith Charitable Trust","doi-asserted-by":"crossref","award":["C2506"],"award-info":[{"award-number":["C2506"]}],"id":[{"id":"10.13039\/100001340","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,6,27]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  RNA splicing is a fundamental feature of eukaryotic gene expression that is co-opted by many nuclear-replicating viruses to produce viral transcripts. Recent clinical development of spliceosome-targeting therapeutics has demonstrated that splicing can be safely modulated\n                  <jats:italic>in vivo,<\/jats:italic>\n                  raising the possibility that viral dependence on host RNA processing may represent an exploitable antiviral vulnerability. Here, we show that pharmacologic inhibition of cellular RNA splicing preferentially suppresses the expression of spliced viral transcripts and profoundly impairs replication of multiple splicing-dependent viruses. In human adenovirus, low-dose inhibition of the spliceosome using mechanistically distinct chemical inhibitors, or genetic depletion of SF3B1, disrupted splicing of complex late viral transcripts while largely sparing simple early transcripts. This resulted in impaired viral DNA replication, reduced late protein accumulation, and multi-log decreases in infectious progeny production. Splicing inhibitors similarly impaired RNA splicing and viral replication of DNA virus Herpes Simplex virus 1 and RNA virus influenza A, which both rely on host-dependent RNA processing strategies. In contrast, cytoplasmic RNA viruses lacking spliceosome dependence were unaffected, supporting an on-target mechanism. Together, these findings identify viral RNA processing complexity as a determinant of antiviral sensitivity and establish host spliceosome dependence as a shared and targetable vulnerability across diverse viral families.\n                <\/jats:p>","DOI":"10.64898\/2026.06.26.734849","type":"posted-content","created":{"date-parts":[[2026,6,28]],"date-time":"2026-06-28T07:35:24Z","timestamp":1782632124000},"source":"Crossref","is-referenced-by-count":0,"title":["Inhibition of RNA splicing is a novel therapeutic strategy for disruption of nuclear replicating viruses"],"prefix":"10.64898","author":[{"given":"Lorenzo","family":"Serra","sequence":"first","affiliation":[{"name":"Genome Regulation and Cell Signaling, Ellen and Ronald Caplan Cancer Center, The Wistar Institute, Philadelphia, PA, United States"},{"name":"Cellular and Molecular Biology Program, Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claire M.","family":"O\u2019Brien","sequence":"additional","affiliation":[{"name":"Genome Regulation and Cell Signaling, Ellen and Ronald Caplan Cancer Center, The Wistar Institute, Philadelphia, PA, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Molly R.","family":"Patterson","sequence":"additional","affiliation":[{"name":"Genome Regulation and Cell Signaling, Ellen and Ronald Caplan Cancer Center, The Wistar Institute, Philadelphia, PA, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clayton J.","family":"Otter","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States"},{"name":"Penn Center for Research on Emerging Viruses, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, Unites States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alison","family":"Yu","sequence":"additional","affiliation":[{"name":"Genome Regulation and Cell Signaling, Ellen and Ronald Caplan Cancer Center, The Wistar Institute, Philadelphia, PA, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reyes W.","family":"Acosta","sequence":"additional","affiliation":[{"name":"Genome Regulation and Cell Signaling, Ellen and Ronald Caplan Cancer Center, The Wistar Institute, Philadelphia, PA, United States"},{"name":"Cell and Molecular Biology Graduate Group, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel T.","family":"Claiborne","sequence":"additional","affiliation":[{"name":"Genome Regulation and Cell Signaling, Ellen and Ronald Caplan Cancer Center, The Wistar Institute, Philadelphia, PA, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susan R.","family":"Weiss","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States"},{"name":"Penn Center for Research on Emerging Viruses, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, Unites States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4654-4604","authenticated-orcid":false,"given":"Alexander M.","family":"Price","sequence":"additional","affiliation":[{"name":"Genome Regulation and Cell Signaling, Ellen and Ronald Caplan Cancer Center, The Wistar Institute, Philadelphia, PA, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026063005400600000_2026.06.26.734849v1.1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-091719-064225"},{"key":"2026063005400600000_2026.06.26.734849v1.2","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a039446"},{"key":"2026063005400600000_2026.06.26.734849v1.3","doi-asserted-by":"crossref","unstructured":"Li, R. et al. Virus usurps alternative splicing to clear the decks for infection. Virol J 20, 131 (2023).","DOI":"10.1186\/s12985-023-02098-9"},{"key":"2026063005400600000_2026.06.26.734849v1.4","doi-asserted-by":"publisher","DOI":"10.1038\/s41429-021-00450-1"},{"key":"2026063005400600000_2026.06.26.734849v1.5","first-page":"1704","volume-title":"Fields Virology","volume":"2","year":"2013"},{"key":"2026063005400600000_2026.06.26.734849v1.6","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.74.8.3171"},{"key":"2026063005400600000_2026.06.26.734849v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(77)90180-5"},{"key":"2026063005400600000_2026.06.26.734849v1.8","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1010797"},{"key":"2026063005400600000_2026.06.26.734849v1.9","first-page":"1823","volume-title":"Fields Virology","volume":"2","year":"2013"},{"key":"2026063005400600000_2026.06.26.734849v1.10","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1007884"},{"key":"2026063005400600000_2026.06.26.734849v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/s44318-025-00398-0"},{"key":"2026063005400600000_2026.06.26.734849v1.12","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00070-14"},{"key":"2026063005400600000_2026.06.26.734849v1.13","doi-asserted-by":"crossref","first-page":"16069","DOI":"10.1038\/nmicrobiol.2016.69","article-title":"Influenza virus mRNA trafficking through host nuclear speckles","volume":"1","year":"2016","journal-title":"Nat Microbiol"},{"key":"2026063005400600000_2026.06.26.734849v1.14","doi-asserted-by":"publisher","DOI":"10.1016\/j.coviro.2022.101254"},{"key":"2026063005400600000_2026.06.26.734849v1.15","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.2007.16"},{"key":"2026063005400600000_2026.06.26.734849v1.16","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.2007.18"},{"key":"2026063005400600000_2026.06.26.734849v1.17","doi-asserted-by":"publisher","DOI":"10.1021\/cb100248e"},{"key":"2026063005400600000_2026.06.26.734849v1.18","doi-asserted-by":"publisher","DOI":"10.1007\/s10637-013-0046-5"},{"key":"2026063005400600000_2026.06.26.734849v1.19","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1182\/blood-2019-123854","article-title":"Results of a Clinical Trial of H3B-8800, a Splicing Modulator, in Patients with Myelodysplastic Syndromes (MDS), Acute Myeloid Leukemia (AML) or Chronic Myelomonocytic Leukemia (CMML)","volume":"134","year":"2019","journal-title":"Blood"},{"key":"2026063005400600000_2026.06.26.734849v1.20","doi-asserted-by":"publisher","DOI":"10.1038\/nm.4493"},{"key":"2026063005400600000_2026.06.26.734849v1.21","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-05204-z"},{"key":"2026063005400600000_2026.06.26.734849v1.22","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkad258"},{"key":"2026063005400600000_2026.06.26.734849v1.23","doi-asserted-by":"crossref","first-page":"e0114625","DOI":"10.1128\/jvi.01146-25","article-title":"Replication-competent adenovirus reporters utilizing endogenous viral expression architecture","volume":"99","year":"2025","journal-title":"J Virol"},{"key":"2026063005400600000_2026.06.26.734849v1.24","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M805556200"},{"key":"2026063005400600000_2026.06.26.734849v1.25","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.3010277"},{"key":"2026063005400600000_2026.06.26.734849v1.26","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccell.2016.08.006"},{"key":"2026063005400600000_2026.06.26.734849v1.27","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms14060"},{"key":"2026063005400600000_2026.06.26.734849v1.28","doi-asserted-by":"publisher","DOI":"10.1023\/A:1010608317361"},{"key":"2026063005400600000_2026.06.26.734849v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.12.031"},{"key":"2026063005400600000_2026.06.26.734849v1.30","doi-asserted-by":"crossref","first-page":"101277","DOI":"10.1016\/j.jbc.2021.101277","article-title":"Modulation of SF3B1 in the pre-mRNA spliceosome induces a RIG-I-dependent type I IFN response","volume":"297","year":"2021","journal-title":"J Biol Chem"},{"key":"2026063005400600000_2026.06.26.734849v1.31","doi-asserted-by":"crossref","first-page":"116384","DOI":"10.1016\/j.celrep.2025.116384","article-title":"Spliceosome inhibition induces Z-RNA and ZBP1-driven cell death in small cell lung cancer","volume":"44","year":"2025","journal-title":"Cell Rep"},{"key":"2026063005400600000_2026.06.26.734849v1.32","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2012.05.005"},{"key":"2026063005400600000_2026.06.26.734849v1.33","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2017.11.008"},{"key":"2026063005400600000_2026.06.26.734849v1.34","doi-asserted-by":"publisher","DOI":"10.1007\/s13365-013-0178-6"},{"key":"2026063005400600000_2026.06.26.734849v1.35","doi-asserted-by":"publisher","DOI":"10.1016\/j.bcp.2022.115322"},{"key":"2026063005400600000_2026.06.26.734849v1.36","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.76.2.841-850.2002"},{"key":"2026063005400600000_2026.06.26.734849v1.37","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.68.12.8118-8124.1994"},{"key":"2026063005400600000_2026.06.26.734849v1.38","doi-asserted-by":"crossref","first-page":"e0175424","DOI":"10.1128\/spectrum.01754-24","article-title":"A drug repurposing screen identifies decitabine as an HSV-1 antiviral","volume":"12","year":"2024","journal-title":"Microbiol Spectr"},{"key":"2026063005400600000_2026.06.26.734849v1.39","doi-asserted-by":"publisher","DOI":"10.4161\/rna.8.2.14513"},{"key":"2026063005400600000_2026.06.26.734849v1.40","doi-asserted-by":"publisher","DOI":"10.1016\/S1097-2765(00)80099-4"},{"key":"2026063005400600000_2026.06.26.734849v1.41","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/17.1.288"},{"key":"2026063005400600000_2026.06.26.734849v1.42","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.67.9.5585-5594.1993"},{"key":"2026063005400600000_2026.06.26.734849v1.43","doi-asserted-by":"publisher","DOI":"10.1016\/j.coviro.2015.01.007"},{"key":"2026063005400600000_2026.06.26.734849v1.44","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2012.12.010"},{"key":"2026063005400600000_2026.06.26.734849v1.45","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002998"},{"key":"2026063005400600000_2026.06.26.734849v1.46","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1186\/s12929-023-00897-4","article-title":"Targeting influenza A virus by splicing inhibitor herboxidiene reveals the importance of subtype-specific signatures around splice sites","volume":"30","year":"2023","journal-title":"J Biomed Sci"},{"key":"2026063005400600000_2026.06.26.734849v1.47","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2024.11.015"},{"key":"2026063005400600000_2026.06.26.734849v1.48","doi-asserted-by":"crossref","first-page":"578","DOI":"10.3390\/v15020578","article-title":"Human Coronavirus OC43 as a Low-Risk Model to Study COVID-19","volume":"15","year":"2023","journal-title":"Viruses"},{"key":"2026063005400600000_2026.06.26.734849v1.49","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1146\/annurev-virology-100114-055218","article-title":"Continuous and Discontinuous RNA Synthesis in Coronaviruses","volume":"2","year":"2015","journal-title":"Annu Rev Virol"},{"key":"2026063005400600000_2026.06.26.734849v1.50","doi-asserted-by":"crossref","first-page":"e0310519","DOI":"10.1371\/journal.pone.0310519","article-title":"Isoginkgetin and Madrasin are poor splicing inhibitors","volume":"19","year":"2024","journal-title":"PLoS One"},{"key":"2026063005400600000_2026.06.26.734849v1.51","doi-asserted-by":"publisher","DOI":"10.7554\/elife.27402"},{"key":"2026063005400600000_2026.06.26.734849v1.52","first-page":"e00489","article-title":"Isoginkgetin, a Natural Biflavonoid Proteasome Inhibitor, Sensitizes Cancer Cells to Apoptosis via Disruption of Lysosomal Homeostasis and Impaired Protein Clearance","volume":"39","year":"2019","journal-title":"Mol Cell Biol"},{"key":"2026063005400600000_2026.06.26.734849v1.53","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2332-7"},{"key":"2026063005400600000_2026.06.26.734849v1.54","doi-asserted-by":"crossref","unstructured":"Xu, J. et al. Influenza neuraminidase mutations and resistance to neuraminidase inhibitors. Emerg Microbes Infect 13, 2429627.","DOI":"10.1080\/22221751.2024.2429627"},{"key":"2026063005400600000_2026.06.26.734849v1.55","doi-asserted-by":"publisher","DOI":"10.1128\/mSystems.00388-21"},{"key":"2026063005400600000_2026.06.26.734849v1.56","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-03511-5"},{"key":"2026063005400600000_2026.06.26.734849v1.57","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00116-13"},{"key":"2026063005400600000_2026.06.26.734849v1.58","doi-asserted-by":"publisher","DOI":"10.1055\/s-0036-1584923"},{"key":"2026063005400600000_2026.06.26.734849v1.59","doi-asserted-by":"publisher","DOI":"10.1016\/j.molmed.2022.10.001"},{"key":"2026063005400600000_2026.06.26.734849v1.60","doi-asserted-by":"crossref","first-page":"105034","DOI":"10.1016\/j.antiviral.2021.105034","article-title":"Emerging antiviral therapeutics for human adenovirus infection: Recent developments and novel strategies","volume":"188","year":"2021","journal-title":"Antiviral Res"},{"key":"2026063005400600000_2026.06.26.734849v1.61","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.virol.2019.09.005","article-title":"Development of a novel screening platform for the identification of small molecule inhibitors of human adenovirus","volume":"538","year":"2019","journal-title":"Virology"},{"key":"2026063005400600000_2026.06.26.734849v1.62","first-page":"e01623","article-title":"Suppression of Adenovirus Replication by Cardiotonic Steroids","volume":"91","year":"2017","journal-title":"J Virol"},{"key":"2026063005400600000_2026.06.26.734849v1.63","doi-asserted-by":"publisher","DOI":"10.1172\/JCI73805"},{"key":"2026063005400600000_2026.06.26.734849v1.64","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1021\/acsinfecdis.7b00009","article-title":"Cell Cycle-Dependent Kinase Cdk9 Is a Postexposure Drug Target against Human Adenoviruses","volume":"3","year":"2017","journal-title":"ACS Infect Dis"},{"key":"2026063005400600000_2026.06.26.734849v1.65","doi-asserted-by":"publisher","DOI":"10.1007\/s00280-002-0527-2"},{"key":"2026063005400600000_2026.06.26.734849v1.66","first-page":"575","article-title":"Administration of 3\u2019-azido-3\u2019-deoxythymidine, an inhibitor of HTLV-III\/LAV replication, to patients with AIDS or AIDS-related complex","volume":"1","year":"1986","journal-title":"Lancet"},{"key":"2026063005400600000_2026.06.26.734849v1.67","doi-asserted-by":"crossref","first-page":"101123","DOI":"10.1016\/j.xpro.2022.101123","article-title":"AdenoBuilder: A platform for the modular assembly of recombinant adenoviruses","volume":"3","year":"2022","journal-title":"STAR Protocols"},{"key":"2026063005400600000_2026.06.26.734849v1.68","doi-asserted-by":"crossref","first-page":"e0215025","DOI":"10.1128\/spectrum.02150-25","article-title":"Bioluminescent reporter influenza A viruses to track viral infections","volume":"13","year":"2025","journal-title":"Microbiol Spectr"},{"key":"2026063005400600000_2026.06.26.734849v1.69","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.76.22.11254-11264.2002"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.06.26.734849","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T13:00:43Z","timestamp":1782824443000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.06.26.734849"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,27]]},"references-count":69,"URL":"https:\/\/doi.org\/10.64898\/2026.06.26.734849","relation":{},"subject":[],"published":{"date-parts":[[2026,6,27]]},"subtype":"preprint"}}