{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:21:45Z","timestamp":1785424905344,"version":"3.56.0"},"reference-count":57,"publisher":"Rockefeller University Press","issue":"7","license":[{"start":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T00:00:00Z","timestamp":1778630400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/rupress.org\/pages\/terms102024"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R35 CA283892"],"award-info":[{"award-number":["R35 CA283892"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P01 CA111412"],"award-info":[{"award-number":["P01 CA111412"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P01 CA065493"],"award-info":[{"award-number":["P01 CA065493"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P30 CA 77598"],"award-info":[{"award-number":["P30 CA 77598"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2026,7,6]]},"abstract":"<jats:p>Natural killer (NK) cells are promising platforms for off-the-shelf immunotherapy, yet nonviral precision engineering remains limited by poor HDR efficiency, DNA toxicity, and manufacturing challenges. The aim of this study was to establish a high-yield, nonviral knock-in platform. Through extensive in-depth rational screens, we achieved \u223c90% HDR insertion of therapeutic payloads while maintaining 100% postediting recovery. By hijacking endogenous transcriptional programs, we installed genetic circuits into defined genomic loci to tune transgene expression. To enable context-dependent therapeutic responses, we integrated a synthetic positive feedback circuit at the CISH locus, which enhanced NK cell persistence and drove strong expression of anti-CD22\/19 dual CAR. A hypoxia-responsive IL-12 circuit gated by the PFKFB4 promoter restored cytotoxicity under environmental stress. Finally, we showed this platform is compatible with GMP manufacturing and supports clinical-scale expansion. These findings provide a scalable framework for programmable, nonviral editing of NK cell effector functions for therapeutic and research applications.<\/jats:p>","DOI":"10.1084\/jem.20260192","type":"journal-article","created":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T06:20:59Z","timestamp":1776925259000},"update-policy":"https:\/\/doi.org\/10.1084\/jem.crossmarkpolicy","source":"Crossref","is-referenced-by-count":0,"title":["Reprogramming endogenous NK circuits by highly efficient nonviral genome editing"],"prefix":"10.1084","volume":"223","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7460-8882","authenticated-orcid":true,"given":"Rih-Sheng","family":"Huang","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Medical School, University of Minnesota 1 Division of Hematology, Oncology and Transplantation, Department of Medicine, , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Masonic Cancer Center, University of Minnesota 2 , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Microbiology, Immunology, and Cancer Biology Graduate Program, Medical School, University of Minnesota 3 , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9700-0151","authenticated-orcid":true,"given":"Shee Kwan","family":"Phung","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Medical School, University of Minnesota 1 Division of Hematology, Oncology and Transplantation, Department of Medicine, , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Masonic Cancer Center, University of Minnesota 2 , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-4940-6110","authenticated-orcid":true,"given":"Darin","family":"Sumstad","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Masonic Cancer Center, University of Minnesota 2 , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Molecular and Cellular Therapeutics, Translational Therapy Shared Resource, University of Minnesota 4 , St. Paul, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-9158-3634","authenticated-orcid":true,"given":"Anna J.","family":"Weis","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Masonic Cancer Center, University of Minnesota 2 , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Medical School, University of Minnesota 5 Gynecologic Oncology, Department of Obstetrics, Gynecology and Women\u2019s Health, , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1471-8895","authenticated-orcid":true,"given":"Quinlan M.","family":"Kile","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Medical School, University of Minnesota 1 Division of Hematology, Oncology and Transplantation, Department of Medicine, , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Masonic Cancer Center, University of Minnesota 2 , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3197-5885","authenticated-orcid":true,"given":"Laura","family":"Bendzick","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Masonic Cancer Center, University of Minnesota 2 , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Medical School, University of Minnesota 5 Gynecologic Oncology, Department of Obstetrics, Gynecology and Women\u2019s Health, , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4207-9151","authenticated-orcid":true,"given":"Melissa J.","family":"Khaw","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Medical School, University of Minnesota 1 Division of Hematology, Oncology and Transplantation, Department of Medicine, , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Masonic Cancer Center, University of Minnesota 2 , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-5074-9564","authenticated-orcid":true,"given":"Young Y.","family":"Vue","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Medical School, University of Minnesota 1 Division of Hematology, Oncology and Transplantation, Department of Medicine, , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Masonic Cancer Center, University of Minnesota 2 , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8734-0996","authenticated-orcid":true,"given":"David H.","family":"McKenna","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Masonic Cancer Center, University of Minnesota 2 , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Molecular and Cellular Therapeutics, Translational Therapy Shared Resource, University of Minnesota 4 , St. Paul, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0239-5440","authenticated-orcid":true,"given":"Philippa R.","family":"Kennedy","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Medical School, University of Minnesota 1 Division of Hematology, Oncology and Transplantation, Department of Medicine, , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Masonic Cancer Center, University of Minnesota 2 , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0339-4944","authenticated-orcid":true,"given":"Jeffrey S.","family":"Miller","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Medical School, University of Minnesota 1 Division of Hematology, Oncology and Transplantation, Department of Medicine, , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Masonic Cancer Center, University of Minnesota 2 , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5945-0634","authenticated-orcid":true,"given":"Martin","family":"Felices","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Medical School, University of Minnesota 1 Division of Hematology, Oncology and Transplantation, Department of Medicine, , Minneapolis, MN,","place":["USA"]},{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"Masonic Cancer Center, University of Minnesota 2 , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2026,5,13]]},"reference":[{"key":"2026051310070445700_bib1","doi-asserted-by":"publisher","first-page":"458","DOI":"10.1038\/s41587-023-01779-8","article-title":"A highly efficient transgene knock-in technology in clinically relevant cell types","volume":"42","author":"Allen","year":"2024","journal-title":"Nat. Biotechnol."},{"key":"2026051310070445700_bib2","doi-asserted-by":"publisher","first-page":"149","DOI":"10.1038\/s41551-023-01073-7","article-title":"Enhancement of the viability of T cells electroporated with DNA via osmotic dampening of the DNA-sensing cGAS\u2013STING pathway","volume":"8","author":"An","year":"2024","journal-title":"Nat. Biomed. Eng."},{"key":"2026051310070445700_bib3","doi-asserted-by":"publisher","first-page":"1987","DOI":"10.1016\/j.ccell.2025.09.013","article-title":"Engineered natural killer cells for cancer therapy","volume":"43","author":"Biederst\u00e4dt","year":"2025","journal-title":"Cancer Cell"},{"key":"2026051310070445700_bib4","doi-asserted-by":"publisher","first-page":"950","DOI":"10.1016\/j.cels.2022.11.005","article-title":"The sound of silence: Transgene silencing in mammalian cell engineering","volume":"13","author":"Cabrera","year":"2022","journal-title":"Cell Syst"},{"key":"2026051310070445700_bib5","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1038\/s41586-025-09212-7","article-title":"Rewiring endogenous genes in CAR T cells for tumour-restricted payload delivery","volume":"644","author":"Chen","year":"2025","journal-title":"Nature"},{"key":"2026051310070445700_bib6","doi-asserted-by":"publisher","first-page":"5697","DOI":"10.1158\/0008-5472.CAN-11-0103","article-title":"IL-12 release by engineered T cells expressing chimeric antigen receptors can effectively muster an antigen-independent macrophage response on tumor cells that have shut down tumor antigen expression","volume":"71","author":"Chmielewski","year":"2011","journal-title":"Cancer Res."},{"key":"2026051310070445700_bib7","doi-asserted-by":"publisher","first-page":"224","DOI":"10.1038\/s41587-019-0032-3","article-title":"CRISPResso2 provides accurate and rapid genome editing sequence analysis","volume":"37","author":"Clement","year":"2019","journal-title":"Nat. Biotechnol."},{"key":"2026051310070445700_bib8","doi-asserted-by":"publisher","first-page":"102157","DOI":"10.1016\/j.xcrm.2025.102157","article-title":"Senescence and inflammation are unintended adverse consequences of CRISPR-Cas9\/AAV6-mediated gene editing in hematopoietic stem cells","volume":"6","author":"Conti","year":"2025","journal-title":"Cell Rep. Med."},{"key":"2026051310070445700_bib9","doi-asserted-by":"publisher","first-page":"1970","DOI":"10.1182\/bloodadvances.2018028332","article-title":"First-in-human trial of rhIL-15 and haploidentical natural killer cell therapy for advanced acute myeloid leukemia","volume":"3","author":"Cooley","year":"2019","journal-title":"Blood Adv."},{"key":"2026051310070445700_bib10","doi-asserted-by":"publisher","first-page":"1778","DOI":"10.1038\/s41587-024-02488-6","article-title":"Genome editing with the HDR-enhancing DNA-PKcs inhibitor AZD7648 causes large-scale genomic alterations","volume":"43","author":"Cullot","year":"2025","journal-title":"Nat. Biotechnol."},{"key":"2026051310070445700_bib11","doi-asserted-by":"publisher","first-page":"624","DOI":"10.1182\/blood.2020007748","article-title":"Targeting a cytokine checkpoint enhances the fitness of armored cord blood CAR-NK cells","volume":"137","author":"Daher","year":"2021","journal-title":"Blood"},{"key":"2026051310070445700_bib12","doi-asserted-by":"publisher","first-page":"816","DOI":"10.1038\/ni.3470","article-title":"CIS is a potent checkpoint in NK cell\u2013mediated tumor immunity","volume":"17","author":"Delconte","year":"2016","journal-title":"Nat. Immunol."},{"key":"2026051310070445700_bib13","doi-asserted-by":"publisher","first-page":"975803","DOI":"10.3389\/fimmu.2022.975803","article-title":"Immunogenicity and toxicity of AAV gene therapy","volume":"13","author":"Ertl","year":"2022","journal-title":"Front. Immunol."},{"key":"2026051310070445700_bib14","doi-asserted-by":"publisher","first-page":"127","DOI":"10.1016\/S0140-6736(24)02462-0","article-title":"Induced pluripotent stem-cell-derived CD19-directed chimeric antigen receptor natural killer cells in B-cell lymphoma: A phase 1, first-in-human trial","volume":"405","author":"Ghobadi","year":"2025","journal-title":"The Lancet"},{"key":"2026051310070445700_bib15","doi-asserted-by":"publisher","DOI":"10.1101\/2025.09.15.676172","article-title":"Repurposing base editors for targeted knock-in and simultaneous knockouts to generate multiplex-edited allogeneic CAR T cells with minimal translocations","author":"Glaser","year":"2025","journal-title":"bioRxiv"},{"key":"2026051310070445700_bib16","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20201529","article-title":"A robust platform for expansion and genome editing of primary human natural killer cells","volume":"218","author":"Huang","year":"2021","journal-title":"J. Exp. Med."},{"key":"2026051310070445700_bib17","doi-asserted-by":"publisher","first-page":"149","DOI":"10.3109\/10425170009033983","article-title":"Cloning and characterization of CIS 1b (cytokine inducible SH2-containing protein lb), an alternative splicing form of CIS 1 gene","volume":"11","author":"Jiang","year":"2000","journal-title":"DNA Seq."},{"key":"2026051310070445700_bib18","doi-asserted-by":"publisher","first-page":"1268","DOI":"10.1016\/j.tibtech.2023.04.002","article-title":"Challenges in scaling up AAV-based gene therapy manufacturing","volume":"41","author":"Jiang","year":"2023","journal-title":"Trends Biotechnol."},{"key":"2026051310070445700_bib19","doi-asserted-by":"publisher","first-page":"100236","DOI":"10.1016\/j.crmeth.2022.100236","article-title":"Optimization and validation of CAR transduction into human primary NK cells using CRISPR and AAV","volume":"2","author":"Kararoudi","year":"2022","journal-title":"Cell Rep. Methods"},{"key":"2026051310070445700_bib20","doi-asserted-by":"publisher","DOI":"10.1101\/2025.09.10.675267","article-title":"Versatile and efficient non-viral integration of large transgenes in human T cells via CRISPR knock-in and engineered integrases","author":"Kassing","year":"2025","journal-title":"bioRxiv"},{"key":"2026051310070445700_bib21","doi-asserted-by":"publisher","first-page":"311","DOI":"10.1016\/j.omtm.2022.03.018","article-title":"Pharmacological interventions enhance virus-free generation of TRAC-replaced CAR T cells","volume":"25","author":"Kath","year":"2022","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"2026051310070445700_bib22","doi-asserted-by":"publisher","first-page":"779","DOI":"10.1016\/j.it.2025.08.007","article-title":"Transcriptional control of natural killer cell antitumor activity","volume":"46","author":"Kee","year":"2025","journal-title":"Trends Immunol."},{"key":"2026051310070445700_bib23","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abc3791","article-title":"Safe harbor-targeted CRISPR-Cas9 homology-independent targeted integration for multimodality reporter gene-based cell tracking","volume":"7","author":"Kelly","year":"2021","journal-title":"Sci. Adv."},{"key":"2026051310070445700_bib24","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adn1849","article-title":"Metabolic programs drive function of therapeutic NK cells in hypoxic tumor environments","volume":"10","author":"Kennedy","year":"2024","journal-title":"Sci. Adv."},{"key":"2026051310070445700_bib25","doi-asserted-by":"publisher","DOI":"10.1136\/jitc-2024-011370","article-title":"Novel trispecific killer engager targeting B7-H3 enhances natural killer cell antitumor activity against head and neck cancer","volume":"13","author":"Khaw","year":"2025","journal-title":"J. Immunother. Cancer"},{"key":"2026051310070445700_bib26","doi-asserted-by":"publisher","first-page":"3422","DOI":"10.1038\/s41467-022-30480-8","article-title":"Cas9-induced large deletions and small indels are controlled in a convergent fashion","volume":"13","author":"Kosicki","year":"2022","journal-title":"Nat. Commun."},{"key":"2026051310070445700_bib27","doi-asserted-by":"publisher","first-page":"2541","DOI":"10.1182\/blood.V90.7.2541","article-title":"Effects of single-dose interleukin-12 exposure on interleukin-12\u2013associated toxicity and interferon-\u03b3 production","volume":"90","author":"Leonard","year":"1997","journal-title":"Blood"},{"key":"2026051310070445700_bib28","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.04766","article-title":"Enhanced homology-directed human genome engineering by controlled timing of CRISPR\/Cas9 delivery","volume":"3","author":"Lin","year":"2014","journal-title":"Elife"},{"key":"2026051310070445700_bib29","doi-asserted-by":"publisher","first-page":"545","DOI":"10.1056\/NEJMoa1910607","article-title":"Use of CAR-transduced natural killer cells in CD19-positive lymphoid tumors","volume":"382","author":"Liu","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"2026051310070445700_bib30","doi-asserted-by":"publisher","first-page":"230","DOI":"10.1093\/stcltm\/szad087","article-title":"Genetic manipulation approaches to enhance the clinical application of NK cell-based immunotherapy","volume":"13","author":"Maia","year":"2024","journal-title":"Stem Cells Transl. Med."},{"key":"2026051310070445700_bib31","doi-asserted-by":"publisher","first-page":"3148","DOI":"10.1182\/blood.V89.9.3148","article-title":"CIS, a cytokine inducible SH2 protein, is a target of the JAK-STAT5 pathway and modulates STAT5 activation","volume":"89","author":"Matsumoto","year":"1997","journal-title":"Blood"},{"key":"2026051310070445700_bib32","doi-asserted-by":"publisher","first-page":"2876","DOI":"10.1038\/s41467-020-16643-5","article-title":"Synergistic gene editing in human iPS cells via cell cycle and DNA repair modulation","volume":"11","author":"Maurissen","year":"2020","journal-title":"Nat. Commun."},{"key":"2026051310070445700_bib33","doi-asserted-by":"publisher","DOI":"10.1136\/jitc-2021-003715","article-title":"T cell receptor engineering of primary NK cells to therapeutically target tumors and tumor immune evasion","volume":"10","author":"Morton","year":"2022","journal-title":"J. Immunother. Cancer"},{"key":"2026051310070445700_bib34","doi-asserted-by":"publisher","first-page":"3059","DOI":"10.1182\/blood.2024027763","article-title":"Chronic NK cell activation results in a dysfunctional, tissue resident\u2013like state mediated by KLF2 deficiency","volume":"146","author":"Myers","year":"2025","journal-title":"Blood"},{"key":"2026051310070445700_bib35","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1038\/s41571-020-0426-7","article-title":"Exploring the NK cell platform for cancer immunotherapy","volume":"18","author":"Myers","year":"2021","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"2026051310070445700_bib36","doi-asserted-by":"publisher","first-page":"44","DOI":"10.1038\/s41587-019-0325-6","article-title":"Polymer-stabilized Cas9 nanoparticles and modified repair templates increase genome editing efficiency","volume":"38","author":"Nguyen","year":"2020","journal-title":"Nat. Biotechnol."},{"key":"2026051310070445700_bib37","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20211530","article-title":"High-efficiency nonviral CRISPR\/Cas9-mediated gene editing of human T cells using plasmid donor DNA","volume":"219","author":"Oh","year":"2022","journal-title":"J. Exp. Med."},{"key":"2026051310070445700_bib38","doi-asserted-by":"publisher","first-page":"2074","DOI":"10.1056\/NEJMoa2401530","article-title":"Indolent CD4+ CAR T-cell lymphoma after cilta-cel CAR T-cell therapy","volume":"390","author":"Ozdemirli","year":"2024","journal-title":"N. Engl. J. Med."},{"key":"2026051310070445700_bib39","doi-asserted-by":"publisher","first-page":"2095","DOI":"10.1084\/jem.20150304","article-title":"Cish actively silences TCR signaling in CD8+ T cells to maintain tumor tolerance","volume":"212","author":"Palmer","year":"2015","journal-title":"J. Exp. Med."},{"key":"2026051310070445700_bib40","doi-asserted-by":"publisher","first-page":"4133","DOI":"10.1182\/blood-2011-12-400044","article-title":"Tumor-targeted T cells modified to secrete IL-12 eradicate systemic tumors without need for prior conditioning","volume":"119","author":"Pegram","year":"2012","journal-title":"Blood"},{"key":"2026051310070445700_bib41","doi-asserted-by":"publisher","first-page":"405","DOI":"10.1038\/s41586-018-0326-5","article-title":"Reprogramming human T cell function and specificity with non-viral genome targeting","volume":"559","author":"Roth","year":"2018","journal-title":"Nature"},{"key":"2026051310070445700_bib42","doi-asserted-by":"publisher","first-page":"1014","DOI":"10.1016\/j.ymthe.2025.01.024","article-title":"Virus-free CRISPR knockin of a chimeric antigen receptor into KLRC1 generates potent GD2-specific natural killer cells","volume":"33","author":"Shankar","year":"2025","journal-title":"Mol. Ther."},{"key":"2026051310070445700_bib43","doi-asserted-by":"publisher","first-page":"1652","DOI":"10.1016\/j.cell.2023.03.006","article-title":"Immune checkpoint therapy\u2014current perspectives and future directions","volume":"186","author":"Sharma","year":"2023","journal-title":"Cell"},{"key":"2026051310070445700_bib44","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20212434","article-title":"Differential IL-12 signaling induces human natural killer cell activating receptor-mediated ligand-specific expansion","volume":"219","author":"Shemesh","year":"2022","journal-title":"J. Exp. Med."},{"key":"2026051310070445700_bib45","doi-asserted-by":"publisher","first-page":"521","DOI":"10.1038\/s41587-022-01418-8","article-title":"High-yield genome engineering in primary cells using a hybrid ssDNA repair template and small-molecule cocktails","volume":"41","author":"Shy","year":"2023","journal-title":"Nat. Biotechnol."},{"key":"2026051310070445700_bib46","doi-asserted-by":"publisher","first-page":"4247","DOI":"10.1182\/blood-2004-11-4564","article-title":"An inducible caspase 9 safety switch for T-cell therapy","volume":"105","author":"Straathof","year":"2005","journal-title":"Blood"},{"key":"2026051310070445700_bib47","doi-asserted-by":"publisher","first-page":"460","DOI":"10.1111\/j.1524-475X.2010.00611.x","article-title":"Importance of housekeeping gene selection for accurate reverse transcription-quantitative polymerase chain reaction in a wound healing model","volume":"18","author":"Turabelidze","year":"2010","journal-title":"Wound Repair Regen."},{"key":"2026051310070445700_bib48","doi-asserted-by":"publisher","first-page":"584","DOI":"10.1056\/NEJMp2400209","article-title":"Secondary cancers after chimeric antigen receptor T-cell therapy","volume":"390","author":"Verdun","year":"2024","journal-title":"N. Engl. J. Med."},{"key":"2026051310070445700_bib49","doi-asserted-by":"publisher","first-page":"711","DOI":"10.1038\/sj.gt.3302213","article-title":"Detection of integration of plasmid DNA into host genomic DNA following intramuscular injection and electroporation","volume":"11","author":"Wang","year":"2004","journal-title":"Gene Ther."},{"key":"2026051310070445700_bib50","doi-asserted-by":"publisher","first-page":"2109","DOI":"10.1038\/s41467-020-15845-1","article-title":"Timed inhibition of CDC7 increases CRISPR-Cas9 mediated templated repair","volume":"11","author":"Wienert","year":"2020","journal-title":"Nat. Commun."},{"key":"2026051310070445700_bib51","doi-asserted-by":"publisher","DOI":"10.1126\/science.adt5199","article-title":"Programmable gene insertion in human cells with a laboratory-evolved CRISPR-associated transposase","volume":"388","author":"Witte","year":"2025","journal-title":"Science"},{"key":"2026051310070445700_bib52","doi-asserted-by":"publisher","first-page":"1821","DOI":"10.1038\/s41587-024-02504-9","article-title":"Efficient non-viral immune cell engineering using circular single-stranded DNA-mediated genomic integration","volume":"43","author":"Xie","year":"2025","journal-title":"Nat. Biotechnol."},{"key":"2026051310070445700_bib53","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-10940-8","article-title":"Armored CAR T cells enhance antitumor efficacy and overcome the tumor microenvironment","volume":"7","author":"Yeku","year":"2017","journal-title":"Sci. Rep."},{"key":"2026051310070445700_bib54","doi-asserted-by":"publisher","first-page":"387","DOI":"10.1038\/s41589-019-0432-1","article-title":"An efficient gene knock-in strategy using 5\u2019-modified double-stranded DNA donors with short homology arms","volume":"16","author":"Yu","year":"2020","journal-title":"Nat. Chem. Biol."},{"key":"2026051310070445700_bib55","doi-asserted-by":"publisher","first-page":"498","DOI":"10.1158\/2326-6066.CIR-15-0231","article-title":"T cells expressing CD19\/CD20 bispecific chimeric antigen receptors prevent antigen escape by malignant B cells","volume":"4","author":"Zah","year":"2016","journal-title":"Cancer Immunol. Res."},{"key":"2026051310070445700_bib56","doi-asserted-by":"publisher","first-page":"216","DOI":"10.1016\/j.omto.2023.08.009","article-title":"Hypoxia-regulated secretion of IL-12 enhances antitumor activity and safety of CD19 CAR-T cells in the treatment of DLBCL","volume":"30","author":"Zhou","year":"2023","journal-title":"Mol. Ther. - Oncolytics."},{"key":"2026051310070445700_bib57","doi-asserted-by":"publisher","first-page":"224","DOI":"10.1016\/j.stem.2020.05.008","article-title":"Metabolic reprograming via deletion of CISH in human iPSC-derived NK cells promotes in vivo persistence and enhances anti-tumor activity","volume":"27","author":"Zhu","year":"2020","journal-title":"Cell Stem Cell"}],"container-title":["Journal of Experimental Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jem\/article-pdf\/doi\/10.1084\/jem.20260192\/2034009\/jem_20260192.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jem\/article-pdf\/doi\/10.1084\/jem.20260192\/2034009\/jem_20260192.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T14:07:13Z","timestamp":1778681233000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jem\/article\/223\/7\/e20260192\/282553\/Reprogramming-endogenous-NK-circuits-by-highly"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,13]]},"references-count":57,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2026,7,6]]}},"URL":"https:\/\/doi.org\/10.1084\/jem.20260192","relation":{},"ISSN":["0022-1007","1540-9538"],"issn-type":[{"value":"0022-1007","type":"print"},{"value":"1540-9538","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,13]]},"article-number":"e20260192"}}