{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T15:54:53Z","timestamp":1772553293600,"version":"3.50.1"},"posted":{"date-parts":[[2026,2,27]]},"group-title":"Molecular Biology","reference-count":44,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T00:00:00Z","timestamp":1772150400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"name":"BMFTR","award":["01KI2105"],"award-info":[{"award-number":["01KI2105"]}]},{"name":"EFRE and IFB"},{"DOI":"10.13039\/100000060","name":"NIAID","doi-asserted-by":"crossref","award":["R01AI181053"],"award-info":[{"award-number":["R01AI181053"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000060","name":"NIAID","doi-asserted-by":"crossref","award":["R01AI145164"],"award-info":[{"award-number":["R01AI145164"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000060","name":"NIAID","doi-asserted-by":"crossref","award":["UM1AI164559"],"award-info":[{"award-number":["UM1AI164559"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000050","name":"NHLBI","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100000050","id-type":"DOI","asserted-by":"crossref"}]},{"name":"NIDA"},{"DOI":"10.13039\/100000025","name":"NIMH","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100000025","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000065","name":"NINDS","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000062","name":"NIDDK","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"crossref"}]},{"name":"LIV"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,2,27]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Recombinant adeno-associated virus (rAAV) vectors are widely used for gene delivery but show limited efficiency in immune cells, including T lymphocytes and natural killer (NK) cells. To overcome this barrier, we developed a modular rAAV vector engineering strategy that integrates capsid retargeting with genome optimization. We report a CD7-targeted rAAV vector (CD7-AAV6\/9) featuring a nanobody-fused hybrid capsid derived from a rationally selected chimeric combination of AAV6 and AAV9. CD7-AAV6\/9 enables efficient and selective transduction of immortalized and primary human T and NK cells\n                  <jats:italic>in vitro<\/jats:italic>\n                  and\n                  <jats:italic>in vivo<\/jats:italic>\n                  in a humanized mouse model, achieves high production titers, and exhibits markedly reduced off-target transduction compared with wild-type serotypes. In parallel, we demonstrate that incorporation of a human gene\u2013derived intron into the vector genome overcomes host-mediated transcriptional repression and enables robust transgene expression in human CD7\u207a T lymphocyte and NK cell populations. To our knowledge, this represents the first application of intron-mediated enhancement in a rAAV vector context. Together, our findings establish an integrated capsid\u2013genome design framework for targeting human T and NK cells, notoriously challenging immune cell populations for gene therapy, and provide a versatile platform readily adaptable to alternative surface markers and therapeutic payloads.\n                <\/jats:p>","DOI":"10.64898\/2026.02.26.707014","type":"posted-content","created":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T23:45:13Z","timestamp":1772235913000},"source":"Crossref","is-referenced-by-count":0,"title":["Development of a recombinant adeno-associated virus vector for human T lymphocyte- and natural killer cell-targeted gene therapy"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-2975-6893","authenticated-orcid":false,"given":"Hendrik","family":"Jahnz","sequence":"first","affiliation":[{"name":"Leibniz Institute of Virology (LIV), Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4089-4227","authenticated-orcid":false,"given":"Martin V.","family":"Hamann","sequence":"additional","affiliation":[{"name":"Leibniz Institute of Virology (LIV), Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongil","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Section of Infectious Diseases, Yale University School of Medicine, New Haven, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yaping","family":"Sun","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Section of Infectious Diseases, Yale University School of Medicine, New Haven, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Natalia Salazar","family":"Quiroz","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Section of Infectious Diseases, Yale University School of Medicine, New Haven, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Li","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Section of Infectious Diseases, Yale University School of Medicine, New Haven, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Umm E","family":"Swaiba","sequence":"additional","affiliation":[{"name":"Leibniz Institute of Virology (LIV), Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel","family":"Foth","sequence":"additional","affiliation":[{"name":"Leibniz Institute of Virology (LIV), Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Niklas","family":"Beschorner","sequence":"additional","affiliation":[{"name":"PROVIREX Genome Editing Therapies GmbH, Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Priti","family":"Kumar","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Section of Infectious Diseases, Yale University School of Medicine, New Haven, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-5274-3108","authenticated-orcid":false,"given":"Ulrike C.","family":"Lange","sequence":"additional","affiliation":[{"name":"Leibniz Institute of Virology (LIV), Hamburg, Germany"},{"name":"Institute for Infection Research and Vaccine Development, University Medical Center Hamburg\u2013Eppendorf, Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2026030304552453000_2026.02.26.707014v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(17)31868-8"},{"key":"2026030304552453000_2026.02.26.707014v1.2","doi-asserted-by":"publisher","DOI":"10.1038\/gt.2012.43"},{"key":"2026030304552453000_2026.02.26.707014v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/S1474-4422(21)00001-6"},{"key":"2026030304552453000_2026.02.26.707014v1.4","doi-asserted-by":"publisher","DOI":"10.1038\/s41392-024-01780-w"},{"key":"2026030304552453000_2026.02.26.707014v1.5","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1407309"},{"key":"2026030304552453000_2026.02.26.707014v1.6","doi-asserted-by":"publisher","DOI":"10.3389\/FMED.2021.809118"},{"key":"2026030304552453000_2026.02.26.707014v1.7","doi-asserted-by":"publisher","DOI":"10.1007\/s40259-017-0234-5"},{"key":"2026030304552453000_2026.02.26.707014v1.8","doi-asserted-by":"publisher","DOI":"10.1089\/HUM.2015.136"},{"key":"2026030304552453000_2026.02.26.707014v1.9","doi-asserted-by":"publisher","DOI":"10.1038\/s41573-019-0012-9"},{"key":"2026030304552453000_2026.02.26.707014v1.10","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2019.264"},{"key":"2026030304552453000_2026.02.26.707014v1.11","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00649-08"},{"key":"2026030304552453000_2026.02.26.707014v1.12","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2017.025"},{"key":"2026030304552453000_2026.02.26.707014v1.13","doi-asserted-by":"publisher","DOI":"10.1186\/1743-422X-10-74"},{"key":"2026030304552453000_2026.02.26.707014v1.14","doi-asserted-by":"publisher","DOI":"10.1016\/j.coviro.2016.08.003"},{"key":"2026030304552453000_2026.02.26.707014v1.15","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2023.10.015"},{"key":"2026030304552453000_2026.02.26.707014v1.16","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0261269"},{"key":"2026030304552453000_2026.02.26.707014v1.17","doi-asserted-by":"publisher","DOI":"10.1182\/blood.v94.3.864.415k34_864_874"},{"key":"2026030304552453000_2026.02.26.707014v1.18","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2024.08.002"},{"key":"2026030304552453000_2026.02.26.707014v1.19","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2023.122399"},{"key":"2026030304552453000_2026.02.26.707014v1.20","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2024.101378"},{"key":"2026030304552453000_2026.02.26.707014v1.21","doi-asserted-by":"publisher","DOI":"10.1093\/narmme\/ugae008"},{"key":"2026030304552453000_2026.02.26.707014v1.22","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2019.09.003"},{"key":"2026030304552453000_2026.02.26.707014v1.23","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00254-08"},{"key":"2026030304552453000_2026.02.26.707014v1.24","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2012.186"},{"key":"2026030304552453000_2026.02.26.707014v1.25","doi-asserted-by":"publisher","DOI":"10.1016\/J.OMTM.2019.01.008"},{"key":"2026030304552453000_2026.02.26.707014v1.26","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.77.20.11072-11081.2003"},{"key":"2026030304552453000_2026.02.26.707014v1.27","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.77.12.6995-7006.2003"},{"key":"2026030304552453000_2026.02.26.707014v1.28","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2008.11.037"},{"key":"2026030304552453000_2026.02.26.707014v1.29","doi-asserted-by":"publisher","DOI":"10.1038\/gt.2015.89"},{"key":"2026030304552453000_2026.02.26.707014v1.30","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-016-0001-8"},{"key":"2026030304552453000_2026.02.26.707014v1.31","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1128\/jvi.68.1.170-176.1994","article-title":"Site-directed mutagenesis of adeno-associated virus type 2 structural protein initiation codons: effects on regulation of synthesis and biological activity","volume":"68","year":"1994","journal-title":"J. Virol"},{"key":"2026030304552453000_2026.02.26.707014v1.32","doi-asserted-by":"publisher","DOI":"10.1038\/mtna.2012.46"},{"key":"2026030304552453000_2026.02.26.707014v1.33","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.799910"},{"key":"2026030304552453000_2026.02.26.707014v1.34","unstructured":"L. Yang , J. Tang , CD7-Nb patent US20170226204A1, 2014."},{"key":"2026030304552453000_2026.02.26.707014v1.35","first-page":"3858","article-title":"Growth of human hepatoma cells lines with differentiated functions in chemically defined medium","volume":"42","year":"1982","journal-title":"Cancer Res"},{"key":"2026030304552453000_2026.02.26.707014v1.36","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-9170-9_8"},{"key":"2026030304552453000_2026.02.26.707014v1.37","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.01827-24"},{"key":"2026030304552453000_2026.02.26.707014v1.38","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2021.04.014"},{"key":"2026030304552453000_2026.02.26.707014v1.39","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-07487-w"},{"key":"2026030304552453000_2026.02.26.707014v1.40","doi-asserted-by":"publisher","DOI":"10.1002\/PRO.4792"},{"key":"2026030304552453000_2026.02.26.707014v1.41","doi-asserted-by":"publisher","DOI":"10.1002\/PRO.3943"},{"key":"2026030304552453000_2026.02.26.707014v1.42","doi-asserted-by":"publisher","DOI":"10.1002\/PRO.3235"},{"key":"2026030304552453000_2026.02.26.707014v1.43","doi-asserted-by":"publisher","DOI":"10.1016\/j.tig.2022.12.005"},{"key":"2026030304552453000_2026.02.26.707014v1.44","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-04228-1"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.02.26.707014","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T12:58:21Z","timestamp":1772542701000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.02.26.707014"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,27]]},"references-count":44,"URL":"https:\/\/doi.org\/10.64898\/2026.02.26.707014","relation":{},"subject":[],"published":{"date-parts":[[2026,2,27]]},"subtype":"preprint"}}