{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T12:23:36Z","timestamp":1789129416120,"version":"build-2803163510"},"reference-count":44,"publisher":"SAGE Publications","issue":"19-20","license":[{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"funder":[{"name":"Lexeo Therapeutics","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Human Gene Therapy"],"published-print":{"date-parts":[[2026,10,1]]},"abstract":"<jats:p>\n                    Adeno-associated virus (AAV) gene transfer vectors mediate long-term expression in nondividing cells, an advantage for treating chronic disorders. However, current platforms lack a way to selectively shut down transgene expression if adverse effects arise. To create an \u201coff switch,\u201d we hypothesized that incorporating unique artificial microRNA (amiRNA) target sequences into an AAV expression cassette would allow subsequent suppression of transgene expression using a second AAV vector encoding the cognate amiRNA. We introduced 22-nt sequences absent from human and mouse transcriptomes into the 3\u2032 untranslated region (UTR) of a therapeutic AAV cassette. To identify optimal amiRNAs, two tandem copies of each amiRNA were cloned into the 3\u2032UTR of an mCherry reporter gene.\n                    <jats:italic toggle=\"yes\">In vitro<\/jats:italic>\n                    assessment of six amiRNA\/target pairs using a dual luciferase assay identified four amiRNAs that efficiently suppressed reporter expression. Cells cotransfected with target site 3 (TS3) and amiRNA-T3B showed the greatest reduction in luciferase activity (80%,\n                    <jats:italic toggle=\"yes\">p<\/jats:italic>\n                    &lt; 0.0001) and were selected for further study. The \u201coff-switch\u201d system was then evaluated using an AAV5 therapeutic vector expressing a recombinant humanized anti-IgE monoclonal antibody (AAV5-TBG-anti-IgE-TS3), designed for long-term suppression of allergen-induced reactions. Co-transfection of HEK293T cells with anti-IgE-TS3 and amiRNA-T3B significantly reduced anti-IgE mRNA and protein levels relative to a control amiRNA (\n                    <jats:italic toggle=\"yes\">p<\/jats:italic>\n                    &lt; 0.0001).\n                    <jats:italic toggle=\"yes\">In vivo<\/jats:italic>\n                    testing in Balb\/c mice (\n                    <jats:italic toggle=\"yes\">n<\/jats:italic>\n                    = 5) involved intravenous administration of AAV5-anti-IgE-TS3 (3.2 \u00d7 10\n                    <jats:sup>10<\/jats:sup>\n                    gc), followed 4 weeks later by an AAVrh.10 amiRNA vector (AAVrh.10-TBG-amiRNA-T3B; 1 \u00d7 10\n                    <jats:sup>11<\/jats:sup>\n                    gc). Control mice receiving only the therapeutic vector expressed 18.4 \u00b1 13.8 \u00b5g\/mL serum anti-IgE at 10 weeks. In contrast, mice receiving the amiRNA \u201coff\u201d vector showed marked suppression of anti-IgE (0.3 \u00b1 0.15 \u00b5g\/mL,\n                    <jats:italic toggle=\"yes\">p<\/jats:italic>\n                    &lt; 0.0001). These findings provide proof-of-concept that AAV-delivered amiRNAs can selectively switch off transgene expression, offering a strategy to improve the safety of AAV-mediated gene therapies.\n                  <\/jats:p>","DOI":"10.1177\/10430342261457812","type":"journal-article","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T07:12:36Z","timestamp":1785913956000},"page":"948-962","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":0,"title":["Suppression of AAV-Delivered Transgene Expression Using Artificial MicroRNAs Delivered by an Alternative AAV Serotype"],"prefix":"10.1177","volume":"37","author":[{"given":"Miguel","family":"de Mulder Rougvie","sequence":"first","affiliation":[{"name":"Department of Genetic Medicine, Weill Cornell Medical College, New York, New York, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mara R.","family":"Kebret","sequence":"additional","affiliation":[{"name":"Department of Genetic Medicine, Weill Cornell Medical College, New York, New York, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youjun","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Genetic Medicine, Weill Cornell Medical College, New York, New York, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katie M.","family":"Stiles","sequence":"additional","affiliation":[{"name":"Department of Genetic Medicine, Weill Cornell Medical College, New York, New York, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6177-6839","authenticated-orcid":false,"given":"Ronald G.","family":"Crystal","sequence":"additional","affiliation":[{"name":"Department of Genetic Medicine, Weill Cornell Medical College, New York, New York, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","published-online":{"date-parts":[[2026,8,5]]},"reference":[{"key":"e_1_3_3_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/gt.2012.6"},{"key":"e_1_3_3_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tips.2021.03.004"},{"key":"e_1_3_3_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.coviro.2017.06.003"},{"key":"e_1_3_3_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41573-019-0012-9"},{"key":"e_1_3_3_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-023-49112-2"},{"key":"e_1_3_3_7_1","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvae207"},{"key":"e_1_3_3_8_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.adm8386"},{"key":"e_1_3_3_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2025.101571"},{"key":"e_1_3_3_10_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.ady6760"},{"key":"e_1_3_3_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41576-019-0205-4"},{"key":"e_1_3_3_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/d41573-021-00017-7"},{"key":"e_1_3_3_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/s40259-017-0234-5"},{"key":"e_1_3_3_14_1","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2018.015"},{"key":"e_1_3_3_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2017.11.007"},{"key":"e_1_3_3_16_1","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2020.29117.trf"},{"key":"e_1_3_3_17_1","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2018.144"},{"key":"e_1_3_3_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2020.08.018"},{"key":"e_1_3_3_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-020-0642-9"},{"key":"e_1_3_3_20_1","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2020.259"},{"key":"e_1_3_3_21_1","doi-asserted-by":"publisher","DOI":"10.1089\/gen.40.09.04"},{"key":"e_1_3_3_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2020.12.007"},{"key":"e_1_3_3_23_1","doi-asserted-by":"publisher","DOI":"10.1002\/advs.202004018"},{"key":"e_1_3_3_24_1","doi-asserted-by":"publisher","DOI":"10.1007\/s40495-016-0073-y"},{"key":"e_1_3_3_25_1","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.aba9188"},{"key":"e_1_3_3_26_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11882-012-0316-x"},{"key":"e_1_3_3_27_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkv1246"},{"key":"e_1_3_3_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2016.03.053"},{"key":"e_1_3_3_29_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkz757"},{"key":"e_1_3_3_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.addr.2007.03.005"},{"key":"e_1_3_3_31_1","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2008.231"},{"key":"e_1_3_3_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtn.2018.12.004"},{"key":"e_1_3_3_33_1","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2016.140"},{"key":"e_1_3_3_34_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2021.669004"},{"key":"e_1_3_3_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtn.2017.12.008"},{"key":"e_1_3_3_36_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0801775105"},{"key":"e_1_3_3_37_1","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2020.00940"},{"key":"e_1_3_3_38_1","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2011.37"},{"key":"e_1_3_3_39_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-019-0357-y"},{"key":"e_1_3_3_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2023.101166"},{"key":"e_1_3_3_41_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.03.023"},{"key":"e_1_3_3_42_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1002693"},{"key":"e_1_3_3_43_1","doi-asserted-by":"publisher","DOI":"10.3390\/ncrna7020027"},{"key":"e_1_3_3_44_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-25078-5"},{"key":"e_1_3_3_45_1","doi-asserted-by":"publisher","DOI":"10.1111\/pbi.13786"}],"container-title":["Human 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