{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,20]],"date-time":"2026-02-20T00:14:53Z","timestamp":1771546493738,"version":"3.50.1"},"reference-count":34,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2020,7,21]],"date-time":"2020-07-21T00:00:00Z","timestamp":1595289600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100010198","name":"Ministerio de Econom\u00eda, Industria y Competitividad, Gobierno de Espa\u00f1a","doi-asserted-by":"publisher","award":["BIO2017\u201389754-C2"],"award-info":[{"award-number":["BIO2017\u201389754-C2"]}],"id":[{"id":"10.13039\/501100010198","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Oncolytic adenoviruses (OAds) present limited efficacy in clinics. The insertion of therapeutic transgenes into OAds genomes, known as \u201carming OAds\u201d, has been the main strategy to improve their therapeutic potential. Different approaches were published in the decade of the 2000s, but with few comparisons. Most armed OAds have complete or partial E3 deletions, leading to a shorter half-life in vivo. We generated E3+ OAds using two insertion sites, After-fiber and After-E4, and two different splice acceptors linked to the major late promoter, either the Ad5 protein IIIa acceptor (IIIaSA) or the Ad40 long fiber acceptor (40SA). The highest transgene levels were obtained with the After-fiber location and 40SA. However, the set of codons of the transgene affected viral fitness, highlighting the relevance of transgene codon usage when arming OAds using the major late promoter.<\/jats:p>","DOI":"10.3390\/ijms21145158","type":"journal-article","created":{"date-parts":[[2020,7,22]],"date-time":"2020-07-22T03:23:16Z","timestamp":1595388196000},"page":"5158","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Arming Oncolytic Adenoviruses: Effect of Insertion Site and Splice Acceptor on Transgene Expression and Viral Fitness"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8170-1276","authenticated-orcid":false,"given":"Mart\u00ed","family":"Farrera-Sal","sequence":"first","affiliation":[{"name":"ProCure Program, Institut Catal\u00e0 d\u2019Oncologia, and Oncobell Program IDIBELL, 08908 L\u2019Hospitalet de Llobregat, Spain"},{"name":"VCN Biosciences S.L., 08174 Sant Cugat, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8563-9273","authenticated-orcid":false,"given":"Jana","family":"de Sostoa","sequence":"additional","affiliation":[{"name":"ProCure Program, Institut Catal\u00e0 d\u2019Oncologia, and Oncobell Program IDIBELL, 08908 L\u2019Hospitalet de Llobregat, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Estela","family":"Nu\u00f1ez-Manch\u00f3n","sequence":"additional","affiliation":[{"name":"Institut d\u2019investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS), Centro de Investigaci\u00f3n Biom\u00e9dica en Red de Enfermedades Raras (CIBERER), Universitat de Barcelona, 08036 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9957-3105","authenticated-orcid":false,"given":"Rafael","family":"Moreno","sequence":"additional","affiliation":[{"name":"ProCure Program, Institut Catal\u00e0 d\u2019Oncologia, and Oncobell Program IDIBELL, 08908 L\u2019Hospitalet de Llobregat, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cristina","family":"Fillat","sequence":"additional","affiliation":[{"name":"Institut d\u2019investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS), Centro de Investigaci\u00f3n Biom\u00e9dica en Red de Enfermedades Raras (CIBERER), Universitat de Barcelona, 08036 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7367-8304","authenticated-orcid":false,"given":"Miriam","family":"Bazan-Peregrino","sequence":"additional","affiliation":[{"name":"VCN Biosciences S.L., 08174 Sant Cugat, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8660-550X","authenticated-orcid":false,"given":"Ramon","family":"Alemany","sequence":"additional","affiliation":[{"name":"ProCure Program, Institut Catal\u00e0 d\u2019Oncologia, and Oncobell Program IDIBELL, 08908 L\u2019Hospitalet de Llobregat, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,7,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1419","DOI":"10.1200\/JCO.2017.75.8219","article-title":"Phase I study of DNX-2401 (Delta-24-RGD) oncolytic adenovirus: Replication and immunotherapeutic effects in recurrent malignant glioma","volume":"36","author":"Lang","year":"2018","journal-title":"J. Clin. Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"10543","DOI":"10.1200\/JCO.2018.36.15_suppl.10543","article-title":"First-in-child trial of celyvir (autologous mesenchymal stem cells carrying the oncolytic virus ICOVIR-5) in patients with relapsed and refractory pediatric solid tumors","volume":"36","author":"Ramirez","year":"2018","journal-title":"J. Clin. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1022","DOI":"10.1038\/sj.cgt.7700542","article-title":"Armed therapeutic viruses: Strategies and challenges to arming oncolytic viruses with therapeutic genes","volume":"9","author":"Hermiston","year":"2002","journal-title":"Cancer Gene Ther."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/0022-2836(92)90922-7","article-title":"Control of adenovirus major late gene expression at multiple levels","volume":"225","author":"Larsson","year":"1992","journal-title":"J. Mol. Boil."},{"key":"ref_5","first-page":"5006","article-title":"Temporal regulation of adenovirus major late alternative RNA splicing","volume":"13","year":"2008","journal-title":"Front. Biosci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5911","DOI":"10.1128\/jvi.67.10.5911-5921.1993","article-title":"Packaging capacity and stability of human adenovirus type 5 vectors","volume":"67","author":"Bett","year":"1993","journal-title":"J. Virol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1128\/jvi.12.3.643-652.1973","article-title":"Use of nondefective adenovirus-simian virus 40 hybrids for mapping the simian virus 40 genome","volume":"12","author":"Kelly","year":"1973","journal-title":"J. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1128\/jvi.16.3.662-673.1975","article-title":"RNA sequences complementary to simian virus 40 and adenovirus 2 DNA in Ad2+NDr- and Ad2+ND3-Infected Cells","volume":"16","author":"Flint","year":"1975","journal-title":"J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"6003","DOI":"10.1093\/nar\/11.17.6003","article-title":"Generation of adenovirus by transfection of plasmids","volume":"11","author":"Berkner","year":"1983","journal-title":"Nucleic Acids Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1328","DOI":"10.1038\/nbt887","article-title":"E3 gene manipulations affect oncolytic adenovirus activity in immunocompetent tumor models","volume":"21","author":"Wang","year":"2003","journal-title":"Nat. Biotechnol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1038\/cgt.2009.12","article-title":"Deletion of the E3-6.7K\/gp19K region reduces the persistence of wild-type adenovirus in a permissive tumor model in Syrian hamsters","volume":"16","author":"Bortolanza","year":"2009","journal-title":"Cancer Gene Ther."},{"key":"ref_12","first-page":"3348","article-title":"The presence of the adenovirus E3 region improves the oncolytic potency of conditionally replicative adenoviruses","volume":"8","author":"Suzuki","year":"2002","journal-title":"Clin. Cancer Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1089\/hum.2010.100","article-title":"Selectivity and efficiency of late transgene expression by transcriptionally targeted oncolytic adenoviruses are dependent on the transgene insertion strategy","volume":"22","author":"Quirin","year":"2011","journal-title":"Hum. Gene Ther."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1038\/sj.gt.3302148","article-title":"5-Fluorocytosine increases the toxicity of Wnt-targeting replicating adenoviruses that express cytosine deaminase as a late gene","volume":"11","author":"Fuerer","year":"2004","journal-title":"Gene Ther."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2317","DOI":"10.1128\/MCB.20.7.2317-2325.2000","article-title":"A novel type of splicing enhancer regulating adenovirus Pre-mRNA splicing","volume":"20","author":"Yue","year":"2000","journal-title":"Mol. Cell. Boil."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1038\/sj.cgt.7700801","article-title":"Tumor cells as cellular vehicles to deliver gene therapies to metastatic tumors","volume":"12","author":"Lillo","year":"2005","journal-title":"Cancer Gene Ther."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1053","DOI":"10.1002\/jgm.754","article-title":"Replication-dependent transgene expression from a conditionally replicating adenovirus via alternative splicing to a heterologous splice-acceptor site","volume":"7","author":"Carette","year":"2005","journal-title":"J. Gene Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1052","DOI":"10.1016\/j.ymthe.2005.07.696","article-title":"Identification of novel insertion sites in the Ad5 genome that utilize the Ad splicing machinery for therapeutic gene expression","volume":"12","author":"Jin","year":"2005","journal-title":"Mol. Ther."},{"key":"ref_19","first-page":"2228","article-title":"Genetic delivery of an immunoRNase by an oncolytic adenovirus enhances anticancer activity","volume":"136","author":"Hammer","year":"2014","journal-title":"Int. J. Cancer"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1038\/sj.cgt.7701093","article-title":"Comparison of the E3 and L3 regions for arming oncolytic adenoviruses to achieve a high level of tumor-specific transgene expression","volume":"15","author":"Robinson","year":"2007","journal-title":"Cancer Gene Ther."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1186\/s40425-019-0505-4","article-title":"Targeting the tumor stroma with an oncolytic adenovirus secreting a fibroblast activation protein-targeted bispecific T-cell engager","volume":"7","author":"Fajardo","year":"2019","journal-title":"J. Immunother. Cancer"},{"key":"ref_22","first-page":"1406","article-title":"Safety and efficacy of VCN-01, an oncolytic adenovirus combining fiber HSG-binding domain replacement with RGD and hyaluronidase expression","volume":"21","author":"Rojas","year":"2014","journal-title":"Clin. Cancer Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1038\/gt.2011.106","article-title":"Improved systemic antitumor therapy with oncolytic adenoviruses by replacing the fiber shaft HSG-binding domain with RGD","volume":"19","author":"Rojas","year":"2011","journal-title":"Gene Ther."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4584","DOI":"10.1158\/1078-0432.CCR-07-5211","article-title":"Effective immunoconjugate therapy in cancer models targeting a serine protease of tumor fibroblasts","volume":"14","author":"Ostermann","year":"2008","journal-title":"Clin. Cancer Res."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"N\u00fa\u00f1ez-Manch\u00f3n, E., Farrera-Sal, M., Castellano, G., Medel, D., Alemany, R., Villanueva, E., and Fillat, C. (2020). Transgene codon usage drives viral fitness and therapeutic efficacy in oncolytic adenoviruses. bioRxiv.","DOI":"10.1101\/2020.06.19.161026"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1038\/sj.gt.3301507","article-title":"Gene delivery from the E3 region of replicating human adenovirus: Evaluation of the 6.7 K\/gp19 K region","volume":"8","author":"Hawkins","year":"2001","journal-title":"Gene Ther."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1142","DOI":"10.1038\/sj.gt.3301509","article-title":"Gene delivery from the E3 region of replicating human adenovirus: Evaluation of the E3B region","volume":"8","author":"Hawkins","year":"2001","journal-title":"Gene Ther."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1132","DOI":"10.1038\/sj.gt.3301508","article-title":"Gene delivery from the E3 region of replicating human adenovirus: Evaluation of the ADP region","volume":"8","author":"Hawkins","year":"2001","journal-title":"Gene Ther."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.ymthe.2005.03.019","article-title":"Development of a transposon-based approach for identifying novel transgene insertion sites within the replicating adenovirus","volume":"12","author":"Kretschmer","year":"2005","journal-title":"Mol. Ther."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1099\/vir.0.83444-0","article-title":"Expression of heterologous genes in oncolytic adenoviruses using picornaviral 2A sequences that trigger ribosome skipping","volume":"89","author":"Funston","year":"2008","journal-title":"J. Gen. Virol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"27546","DOI":"10.1038\/srep27546","article-title":"Codon optimization of the adenoviral fiber negatively impacts structural protein expression and viral fitness","volume":"6","author":"Villanueva","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"659","DOI":"10.2144\/000112993","article-title":"Re-engineering adenovirus vector systems to enable high-throughput analyses of gene function","volume":"45","author":"Stanton","year":"2008","journal-title":"BioTechniques"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1038\/gt.2014.52","article-title":"iRGD tumor-penetrating peptide-modified oncolytic adenovirus shows enhanced tumor transduction, intratumoral dissemination and antitumor efficacy","volume":"21","author":"Rojas","year":"2014","journal-title":"Gene Ther."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"8928","DOI":"10.1158\/0008-5472.CAN-08-1145","article-title":"Bioselection of a gain of function mutation that enhances adenovirus 5 release and improves its antitumoral potency","volume":"68","author":"Gros","year":"2008","journal-title":"Cancer Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/14\/5158\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:50:22Z","timestamp":1760176222000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/14\/5158"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,21]]},"references-count":34,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2020,7]]}},"alternative-id":["ijms21145158"],"URL":"https:\/\/doi.org\/10.3390\/ijms21145158","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,7,21]]}}}