{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,6]],"date-time":"2026-11-06T20:29:39Z","timestamp":1793996979095,"version":"4.8.0"},"reference-count":122,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T00:00:00Z","timestamp":1764892800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>\n                    CRISPR-based gene editing has become a transformative tool to enhance immune cell therapies. In particular, engineering natural killer (NK) cells with CRISPR\/Cas systems has gained traction due to their ability to mediate strong anti-tumor responses in an MHC-unrestricted, non-alloreactive manner. Early trials show the feasibility and safety of allogeneic NK cells, paving the way as scalable \u201coff-the-shelf\u201d products. CRISPR\/Cas9 edits genomes by inducing DNA double-strand breaks (DSBs), mainly repaired through non-homologous end joining (NHEJ) or homology-directed repair (HDR). While effective, CRISPR carries risks of off-target (OT) activity that may disrupt essential genes, cause chromosomal rearrangements, or trigger oncogenic changes - posing threats to product integrity and patient safety. These concerns intensify with multiplex editing, where multiple loci are modified to improve function, persistence, and immune evasion. Since unmodified NK cells are typically short-lived, many clinical-stage products are engineered to express IL-15 or related constructs, extending their half-life and amplifying risks associated with unintended changes. This underscores the urgent need for robust safety assessments. In this review, we summarize the current landscape of safety assessment platforms for evaluating gene edited NK cells. We highlight predictive in silico tools, biochemical\n                    <jats:italic>in vitro<\/jats:italic>\n                    assays, and emerging cell-based detection systems to identify and quantify CRISPR-induced OT events. Particular attention is given to their suitability, limitations, and practical use in primary NK cells and multiplex editing strategies. Our aim is to support the design of safe, effective editing workflows for NK cell therapies - ensuring rigor as the field advances rapidly toward clinical application.\n                  <\/jats:p>","DOI":"10.3389\/fimmu.2025.1711414","type":"journal-article","created":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T06:28:53Z","timestamp":1764916133000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":5,"title":["Engineering with care: safety assessment platforms for CRISPR-modified natural killer cells"],"prefix":"10.3389","volume":"16","author":[{"given":"Alieh","family":"Fazeli","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evelyn","family":"Ullrich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toni","family":"Cathomen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tobias","family":"Bexte","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,12,5]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1225829","article-title":"A programmable dual-RNA\u2013guided DNA endonuclease in adaptive bacterial immunity","volume":"337","author":"Jinek","year":"2012","journal-title":"science"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1208507109","article-title":"Cas9\u2013crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria","volume":"109","author":"Gasiunas","year":"2012","journal-title":"Proc Natl Acad Sci"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-1978-5","article-title":"The promise and challenge of therapeutic genome editing","volume":"578","author":"Doudna","year":"2020","journal-title":"Nature"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-020-0840-5","article-title":"Safety and feasibility of CRISPR-edited T cells in patients with refractory non-small-cell lung cancer","volume":"26","author":"Lu","year":"2020","journal-title":"Nat Med"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2031054","article-title":"CRISPR-Cas9 gene editing for sickle cell disease and \u03b2-thalassemia","volume":"384","author":"Frangoul","year":"2021","journal-title":"New Engl J Med"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-022-01906-z","article-title":"CRISPR\u2013Cas9-mediated gene editing of the BCL11A enhancer for pediatric \u03b20\/\u03b20 transfusion-dependent \u03b2-thalassemia","volume":"28","author":"Fu","year":"2022","journal-title":"Nat Med"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1126\/science.1231143","article-title":"Multiplex genome engineering using CRISPR\/Cas systems","volume":"339","author":"Cong","year":"2013","journal-title":"Science"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1126\/science.1232033","article-title":"RNA-guided human genome engineering via Cas9","volume":"339","author":"Mali","year":"2013","journal-title":"Science"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2507","article-title":"Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease","volume":"31","author":"Cho","year":"2013","journal-title":"Nat Biotechnol"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.00471","article-title":"RNA-programmed genome editing in human cells","volume":"2","author":"Jinek","year":"2013","journal-title":"elife"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-023-06945-1","article-title":"Natural killer cell therapies","volume":"626","author":"Vivier","year":"2024","journal-title":"Nature"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1038\/s41568-022-00491-0","article-title":"Natural killer cells in antitumour adoptive cell immunotherapy","volume":"22","author":"Laskowski","year":"2022","journal-title":"Nat Rev Cancer"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20231222","article-title":"Five decades of natural killer cell discovery","volume":"221","author":"Lanier","year":"2024","journal-title":"J Exp Med"},{"key":"B14","doi-asserted-by":"publisher","first-page":"1481","DOI":"10.3390\/cancers13061481","article-title":"Arming immune cells for battle: a brief journey through the advancements of T and NK cell immunotherapy","volume":"13","author":"Wendel","year":"2021","journal-title":"Cancers"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-023-02785-8","article-title":"Safety, efficacy and determinants of response of allogeneic CD19-specific CAR-NK cells in CD19+ B cell tumors: a phase 1\/2 trial","volume":"30","author":"Marin","year":"2024","journal-title":"Nat Med"},{"key":"B16","doi-asserted-by":"publisher","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":"New Engl J Med"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1007\/s12185-021-03209-4","article-title":"Engineering the next generation of CAR-NK immunotherapies","volume":"114","author":"Biederst\u00e4dt","year":"2021","journal-title":"Int J hematol"},{"key":"B18","doi-asserted-by":"publisher","first-page":"46","DOI":"10.1186\/s40164-025-00633-8","article-title":"The clinical landscape of CAR NK cells","volume":"14","author":"J\u00f8rgensen","year":"2025","journal-title":"Exp Hematol Oncol"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.1063303","article-title":"Multiplexed engineering and precision gene editing in cellular immunotherapy","volume":"13","author":"Biederst\u00e4dt","year":"2022","journal-title":"Front Immunol"},{"key":"B20","doi-asserted-by":"publisher","first-page":"1777651","DOI":"10.1080\/2162402X.2020.1777651","article-title":"Immunotherapy with NK cells: recent developments in gene modification open up new avenues","volume":"9","author":"Reindl","year":"2020","journal-title":"Oncoimmunology"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.3343\/alm.2024.0380","article-title":"Advancing natural killer cell therapy: genetic engineering strategies for enhanced cancer immunotherapy","volume":"45","author":"Park","year":"2025","journal-title":"Ann Lab Med"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1093\/jleuko\/qiad074","article-title":"Nonviral technologies can pave the way for CAR-NK cell therapy","volume":"114","author":"Bexte","year":"2023","journal-title":"J Leukocyte Biol"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.01008","article-title":"Enhanced NK-92 cytotoxicity by CRISPR genome engineering using Cas9 ribonucleoproteins","volume":"11","author":"Huang","year":"2020","journal-title":"Front Immunol"},{"key":"B24","doi-asserted-by":"publisher","first-page":"2081415","DOI":"10.1080\/2162402X.2022.2081415","article-title":"CRISPR-Cas9 based gene editing of the immune checkpoint NKG2A enhances NK cell mediated cytotoxicity against multiple myeloma","volume":"11","author":"Bexte","year":"2022","journal-title":"Oncoimmunology"},{"key":"B25","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":"B26","doi-asserted-by":"publisher","first-page":"406934","DOI":"10.1101\/406934","article-title":"Efficient genome editing of human natural killer cells by CRISPR RNP","author":"Rautela","year":"2018","journal-title":"BioRxiv"},{"key":"B27","doi-asserted-by":"publisher","first-page":"3237","DOI":"10.1182\/blood-2019-129162","article-title":"Primary human NK cell gene-editing reveals a critical role for NKG2A in cytokine-induced memory-like NK cell responses","volume":"134","author":"Berrien-Elliott","year":"2019","journal-title":"Blood"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-024-02282-4","article-title":"In vivo AAV\u2013SB-CRISPR screens of tumor-infiltrating primary NK cells identify genetic checkpoints of CAR-NK therapy","volume":"43","author":"Peng","year":"2024","journal-title":"Nat Biotechnol"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.712722","article-title":"Manipulating the NKG2D receptor-ligand axis using CRISPR: novel technologies for improved host immunity","volume":"12","author":"Alves","year":"2021","journal-title":"Front Immunol"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1002\/JLB.2MA0620-074R","article-title":"Drug target validation in primary human natural killer cells using CRISPR RNP","volume":"108","author":"Rautela","year":"2020","journal-title":"J Leucocyte Biol"},{"key":"B31","doi-asserted-by":"publisher","first-page":"1271_4180","DOI":"10.4049\/jimmunol.212.supp.1271.4180","article-title":"Loss of Siglec-7 drives enhanced NK cell function and correlates with protection from malaria symptoms","volume":"212","author":"Dick","year":"2024","journal-title":"J Immunol"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1182\/bloodadvances.2021006805","article-title":"Targeting hypersialylation in multiple myeloma represents a novel approach to enhance NK cell\u2013mediated tumor responses","volume":"6","author":"Daly","year":"2022","journal-title":"Blood Advances"},{"key":"B33","doi-asserted-by":"publisher","first-page":"52","DOI":"10.1016\/j.ymthe.2019.10.009","article-title":"A genetically engineered primary human natural killer cell platform for cancer immunotherapy","volume":"28","author":"Pomeroy","year":"2020","journal-title":"Mol Ther"},{"key":"B34","doi-asserted-by":"publisher","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 J Am Soc Hematol"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1158\/1538-7445.AM2024-1328","article-title":"AccuBaseTM (ceBE), a second-generation high-fidelity base editor, can proficiently modify genes in a human primary NK cells with minimizing off-target effect","volume":"84","author":"Ying","year":"2024","journal-title":"Cancer Res"},{"key":"B36","doi-asserted-by":"publisher","first-page":"12828","DOI":"10.3390\/ijms232112828","article-title":"Preclinical evaluation of CRISPR-edited CAR-NK-92 cells for off-the-shelf treatment of AML and B-ALL","volume":"23","author":"Ure\u00f1a-Bail\u00e9n","year":"2022","journal-title":"Int J Mol Sci"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1136\/jitc-2024-009560","article-title":"Precision enhancement of CAR-NK cells through non-viral engineering and highly multiplexed base editing","volume":"13","author":"Wang","year":"2025","journal-title":"J ImmunoTher Cancer"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.2200488","article-title":"Ablation of SYK kinase from expanded primary human NK cells via CRISPR\/Cas9 enhances cytotoxicity and cytokine production","volume":"210","author":"Dahlvang","year":"2023","journal-title":"J Immunol"},{"key":"B39","doi-asserted-by":"publisher","first-page":"58237","DOI":"10.3791\/58237","article-title":"Generation of knock-out primary and expanded human NK cells using Cas9 ribonucleoproteins","author":"Kararoudi","year":"2018","journal-title":"J visualized experiments: JoVE"},{"key":"B40","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1182\/blood-2020-139988","article-title":"Preclinical development of edit-201, a multigene edited healthy donor nk cell with enhanced anti-tumor function and superior serial killing activity in an immunosuppressive environment","volume":"136","author":"Borges","year":"2020","journal-title":"Blood"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1182\/blood.2023020973","article-title":"Integration of \u03b6-deficient CARs into the CD3 \u03b6 gene conveys potent cytotoxicity in T and NK cells","volume":"143","author":"Kath","year":"2024","journal-title":"Blood"},{"key":"B42","doi-asserted-by":"publisher","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":"B43","doi-asserted-by":"publisher","first-page":"8439","DOI":"10.1038\/s41467-024-52388-1","article-title":"CRISPR\/Cas9 editing of NKG2A improves the efficacy of primary CD33-directed chimeric antigen receptor natural killer cells","volume":"15","author":"Bexte","year":"2024","journal-title":"Nat Commun"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2022.08.004","article-title":"Harnessing IL-15 signaling to potentiate NK cell-mediated cancer immunotherapy","volume":"43","author":"Ma","year":"2022","journal-title":"Trends Immunol"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1038\/leu.2017.226","article-title":"Cord blood NK cells engineered to express IL-15 and a CD19-targeted CAR show long-term persistence and potent antitumor activity","volume":"32","author":"Liu","year":"2018","journal-title":"Leukemia"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-025-09087-8","article-title":"CREM is a regulatory checkpoint of CAR and IL-15 signalling in NK cells","volume":"643","author":"Rafei","year":"2025","journal-title":"Nature"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.2024.42.16_suppl.TPS5626","article-title":"Phase I\/II study of TROP2 CAR engineered IL15-transduced cord blood-derived NK cells delivered intraperitoneally for the management of platinum resistant ovarian cancer, mesonephric-like adenocarcinoma, and pancreatic cancer","volume":"42","author":"Knisely","year":"2024","journal-title":"Am Soc Clin Oncol"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.add6997","article-title":"Loss of metabolic fitness drives tumor resistance after CAR-NK cell therapy and can be overcome by cytokine engineering","volume":"9","author":"Li","year":"2023","journal-title":"Sci Adv"},{"key":"B49","doi-asserted-by":"publisher","first-page":"1450","DOI":"10.1016\/j.ccell.2024.07.007","article-title":"Interleukin-21 engineering enhances NK cell activity against glioblastoma via CEBPD","volume":"42","author":"Shanley","year":"2024","journal-title":"Cancer Cell"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.1136\/jitc-2021-003894","article-title":"Engineering CAR-NK cells to secrete IL-15 sustains their anti-AML functionality but is associated with systemic toxicities","volume":"9","author":"Christodoulou","year":"2021","journal-title":"J immunother cancer"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3609","article-title":"Genome-wide analysis reveals specificities of Cpf1 endonucleases in human cells","volume":"34","author":"Kim","year":"2016","journal-title":"Nat Biotechnol"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.4192","article-title":"Repair of double-strand breaks induced by CRISPR\u2013Cas9 leads to large deletions and complex rearrangements","volume":"36","author":"Kosicki","year":"2018","journal-title":"Nat Biotechnol"},{"key":"B53","doi-asserted-by":"publisher","first-page":"1136","DOI":"10.1038\/s41467-019-09006-2","article-title":"CRISPR-Cas9 genome editing induces megabase-scale chromosomal truncations","volume":"10","author":"Cullot","year":"2019","journal-title":"Nat Commun"},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2623","article-title":"High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells","volume":"31","author":"Fu","year":"2013","journal-title":"Nat Biotechnol"},{"key":"B55","doi-asserted-by":"publisher","DOI":"10.1007\/s13238-015-0153-5","article-title":"CRISPR\/Cas9-mediated gene editing in human tripronuclear zygotes","volume":"6","author":"Liang","year":"2015","journal-title":"Protein Cell"},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-018-0049-z","article-title":"CRISPR\u2013Cas9 genome editing induces a p53-mediated DNA damage response","volume":"24","author":"Haapaniemi","year":"2018","journal-title":"Nat Med"},{"key":"B57","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-018-0050-6","article-title":"p53 inhibits CRISPR\u2013Cas9 engineering in human pluripotent stem cells","volume":"24","author":"Ihry","year":"2018","journal-title":"Nat Med"},{"key":"B58","doi-asserted-by":"publisher","DOI":"10.1038\/mtna.2015.37","article-title":"Off-target effects in CRISPR\/Cas9-mediated genome engineering","volume":"4","author":"Zhang","year":"2015","journal-title":"Mol Ther Nucleic Acids"},{"key":"B59","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-020-0561-9","article-title":"Genome editing with CRISPR\u2013Cas nucleases, base editors, transposases and prime editors","volume":"38","author":"Anzalone","year":"2020","journal-title":"Nat Biotechnol"},{"key":"B60","doi-asserted-by":"publisher","DOI":"10.1136\/jitc-2025-012841","article-title":"Next-generation multiplex-edited CAR-NK cells: more edits, more power","volume":"13","author":"Bexte","year":"2025","journal-title":"J Immunother Cancer"},{"key":"B61","article-title":"Evaluate CRISPR-Cas9 edits quickly and accurately with rhAmpSeq targeted sequencing","author":"Schubert","year":"2024"},{"key":"B62","doi-asserted-by":"publisher","DOI":"10.1089\/gen.41.09.24","article-title":"CRISPR Applications in Medicine Depend on Minimizing Off-Target Editing: Integrated DNA Technologies describes methods that can be used to quantify and characterize off-target editing in gene editing experiments","volume":"41","author":"Turk","year":"2021","journal-title":"Genet Eng Biotechnol News"},{"key":"B63","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2016.07.004","article-title":"Methods for optimizing CRISPR-Cas9 genome editing specificity","volume":"63","author":"Tycko","year":"2016","journal-title":"Mol Cell"},{"key":"B64","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-021-00525-1","article-title":"Publisher Correction: Precision genome editing using cytosine and adenine base editors in mammalian cells","volume":"16","author":"Huang","year":"2021","journal-title":"Nat Protoc"},{"key":"B65","volume-title":"Human Gene Therapy Products Incorporating Human Genome Editing: Draft Guidance for Industry","author":"Harris","year":"2022"},{"key":"B66","doi-asserted-by":"publisher","DOI":"10.1038\/d41573-024-00050-2","article-title":"Genome-editing medicinal products: the EMA perspective","volume":"23","author":"Tavridou","year":"2024","journal-title":"Nat Rev Drug Discov"},{"key":"B67","doi-asserted-by":"publisher","first-page":"148","DOI":"10.1186\/s13059-016-1012-2","article-title":"Evaluation of off-target and on-target scoring algorithms and integration into the guide RNA selection tool CRISPOR","volume":"17","author":"Haeussler","year":"2016","journal-title":"Genome Biol"},{"key":"B68","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0124633","article-title":"CCTop: an intuitive, flexible and reliable CRISPR\/Cas9 target prediction tool","volume":"10","author":"Stemmer","year":"2015","journal-title":"PloS One"},{"key":"B69","doi-asserted-by":"publisher","DOI":"10.1038\/mtna.2014.64","article-title":"COSMID: a web-based tool for identifying and validating CRISPR\/Cas off-target sites","volume":"3","author":"Cradick","year":"2014","journal-title":"Mol Ther Nucleic Acids"},{"key":"B70","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt1326","article-title":"An online bioinformatics tool predicts zinc finger and TALE nuclease off-target cleavage","volume":"42","author":"Fine","year":"2014","journal-title":"Nucleic Acids Res"},{"key":"B71","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.4061","article-title":"Deep learning improves prediction of CRISPR\u2013Cpf1 guide RNA activity","volume":"36","author":"Kim","year":"2018","journal-title":"Nat Biotechnol"},{"key":"B72","doi-asserted-by":"publisher","first-page":"542","DOI":"10.1186\/s12859-018-2585-4","article-title":"Web-based design and analysis tools for CRISPR base editing","volume":"19","author":"Hwang","year":"2018","journal-title":"BMC Bioinf"},{"key":"B73","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1038\/s41408-022-00660-2","article-title":"Primary CD33-targeting CAR-NK cells for the treatment of acute myeloid leukemia","volume":"12","author":"Albinger","year":"2022","journal-title":"Blood Cancer J"},{"key":"B74","doi-asserted-by":"publisher","DOI":"10.1158\/2159-8290.CD-20-0312","article-title":"Multidimensional analyses of donor memory-like NK cells reveal new associations with response after adoptive immunotherapy for leukemia","volume":"10","author":"Berrien-Elliott","year":"2020","journal-title":"Cancer discov"},{"key":"B75","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-018-0055-0","article-title":"Defining CRISPR\u2013Cas9 genome-wide nuclease activities with CIRCLE-seq","volume":"13","author":"Lazzarotto","year":"2018","journal-title":"Nat Protoc"},{"key":"B76","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4284","article-title":"Mapping the genomic landscape of CRISPR\u2013Cas9 cleavage","volume":"14","author":"Cameron","year":"2017","journal-title":"Nat Methods"},{"key":"B77","doi-asserted-by":"publisher","DOI":"10.1101\/gr.236620.118","article-title":"DIG-seq: a genome-wide CRISPR off-target profiling method using chromatin DNA","volume":"28","author":"Kim","year":"2018","journal-title":"Genome Res"},{"key":"B78","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-020-0555-7","article-title":"CHANGE-seq reveals genetic and epigenetic effects on CRISPR\u2013Cas9 genome-wide activity","volume":"38","author":"Lazzarotto","year":"2020","journal-title":"Nat Biotechnol"},{"key":"B79","doi-asserted-by":"publisher","DOI":"10.1101\/2024.03.28.586621","article-title":"CHANGE-seq-BE enables simultaneously sensitive and unbiased in vitro profiling of base editor genome-wide activity","author":"Lazzarotto","year":"2024","journal-title":"bioRxiv"},{"key":"B80","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3117","article-title":"GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases","volume":"33","author":"Tsai","year":"2015","journal-title":"Nat Biotechnol"},{"key":"B81","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-023-01792-x","article-title":"Deep learning models to predict the editing efficiencies and outcomes of diverse base editors","volume":"42","author":"Kim","year":"2024","journal-title":"Nat Biotechnol"},{"key":"B82","doi-asserted-by":"publisher","first-page":"17231","DOI":"10.3390\/ijms242417231","article-title":"Nicotinamide-expanded allogeneic natural killer cells with CD38 deletion, expressing an enhanced CD38 chimeric antigen receptor, target multiple myeloma cells","volume":"24","author":"Edri","year":"2023","journal-title":"Int J Mol Sci"},{"key":"B83","doi-asserted-by":"publisher","first-page":"437","DOI":"10.3324\/haematol.2020.271908","article-title":"CD38 knockout natural killer cells expressing an affinity optimized CD38 chimeric antigen receptor successfully target acute myeloid leukemia with reduced effector cell fratricide","volume":"107","author":"Gurney","year":"2020","journal-title":"Haematologica"},{"key":"B84","doi-asserted-by":"publisher","DOI":"10.17504\/protocols.io.uvuew6w","article-title":"DISCOVER-Seq: MRE11 ChIP Seq","author":"Wienert","year":"2018"},{"key":"B85","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2408","article-title":"Nucleotide-resolution DNA double-strand break mapping by next-generation sequencing","volume":"10","author":"Crosetto","year":"2013","journal-title":"Nat Methods"},{"key":"B86","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3101","article-title":"Genome-wide detection of DNA double-stranded breaks induced by engineered nucleases","volume":"33","author":"Frock","year":"2015","journal-title":"Nat Biotechnol"},{"key":"B87","doi-asserted-by":"publisher","DOI":"10.3389\/fcell.2021.718466","article-title":"The off-targets of clustered regularly interspaced short palindromic repeats gene editing","volume":"9","author":"Vicente","year":"2021","journal-title":"Front Cell Dev Biol"},{"key":"B88","doi-asserted-by":"publisher","first-page":"1136","DOI":"10.1016\/j.stem.2021.02.002","article-title":"Quantitative evaluation of chromosomal rearrangements in gene-edited human stem cells by CAST-Seq","volume":"28","author":"Turchiano","year":"2021","journal-title":"Cell Stem Cell"},{"key":"B89","doi-asserted-by":"publisher","DOI":"10.3389\/fgeed.2023.1130736","article-title":"T-CAST: An optimized CAST-Seq pipeline for TALEN confirms superior safety and efficacy of obligate-heterodimeric scaffolds","volume":"5","author":"Rhiel","year":"2023","journal-title":"Front Genome Editing"},{"key":"B90","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2024.03.006","article-title":"On-and off-target effects of paired CRISPR-Cas nickase in primary human cells","volume":"32","author":"Klermund","year":"2024","journal-title":"Mol Ther"},{"key":"B91","doi-asserted-by":"publisher","first-page":"243","DOI":"10.1016\/j.jaci.2023.08.003","article-title":"Gene editing of hematopoietic stem cells restores T-cell response in familial hemophagocytic lymphohistiocytosis","volume":"153","author":"Dettmer-Monaco","year":"2024","journal-title":"J Allergy Clin Immunol"},{"key":"B92","doi-asserted-by":"publisher","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":"2024","journal-title":"Nat Biotechnol"},{"key":"B93","doi-asserted-by":"publisher","first-page":"1291","DOI":"10.1038\/s42003-024-06959-z","article-title":"Gene editing of NCF1 loci is associated with homologous recombination and chromosomal rearrangements","volume":"7","author":"Raimondi","year":"2024","journal-title":"Commun Biol"},{"key":"B94","doi-asserted-by":"publisher","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":"B95","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.aan0820","article-title":"Preclinical modeling highlights the therapeutic potential of hematopoietic stem cell gene editing for correction of SCID-X1","volume":"9","author":"Schiroli","year":"2017","journal-title":"Sci Trans Med"},{"key":"B96","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2023.02.011","article-title":"Comparative analysis of CRISPR off-target discovery tools following ex vivo editing of CD34+ hematopoietic stem and progenitor cells","volume":"31","author":"Cromer","year":"2023","journal-title":"Mol Ther"},{"key":"B97","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtn.2025.102636","article-title":"Off-target effects in CRISPR-Cas genome editing for human therapeutics: Progress and challenges","volume":"36","author":"Kalter","year":"2025","journal-title":"Mol Ther Nucleic Acids"},{"key":"B98","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2022.834002","article-title":"CRISPR gene editing of human primary NK and T cells for cancer immunotherapy","volume":"12","author":"Elmas","year":"2022","journal-title":"Front Oncol"},{"key":"B99","doi-asserted-by":"publisher","first-page":"4","DOI":"10.1186\/s13059-022-02842-4","article-title":"Extru-seq: a method for predicting genome-wide Cas9 off-target sites with advantages of both cell-based and in vitro approaches","volume":"24","author":"Kwon","year":"2023","journal-title":"Genome Biol"},{"key":"B100","doi-asserted-by":"publisher","first-page":"598","DOI":"10.4049\/immunohorizons.2400059","article-title":"Chick embryo chorioallantoic membrane as a platform for assessing the in vivo efficacy of chimeric antigen receptor T-cell therapy in solid tumors","volume":"8","author":"Nipper","year":"2024","journal-title":"ImmunoHorizons"},{"key":"B101","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2022.945149","article-title":"A multi-organ-on-chip to recapitulate the infiltration and the cytotoxic activity of circulating NK cells in 3D matrix-based tumor model","volume":"10","author":"Marzagalli","year":"2022","journal-title":"Front bioengineering Biotechnol"},{"key":"B102","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1254821","article-title":"Preclinical assessment of CAR-NK cell-mediated killing efficacy and pharmacokinetics in a rapid zebrafish xenograft model of metastatic breast cancer","volume":"14","author":"Murali Shankar","year":"2023","journal-title":"Front Immunol"},{"key":"B103","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1209237","article-title":"Murine models to study human NK cells in human solid tumors","volume":"14","author":"Parodi","year":"2023","journal-title":"Front Immunol"},{"key":"B104","doi-asserted-by":"publisher","DOI":"10.1007\/s43188-022-00138-0","article-title":"A general toxicity and biodistribution study of human natural killer cells by single or repeated intravenous dose in severe combined immune deficient mice","volume":"38","author":"Park","year":"2022","journal-title":"Toxicological Res"},{"key":"B105","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.992232","article-title":"Preclinical and clinical studies of CAR-NK-cell therapies for Malignancies","volume":"13","author":"Li","year":"2022","journal-title":"Front Immunol"},{"key":"B106","doi-asserted-by":"publisher","DOI":"10.26508\/lsa.201800195","article-title":"Human NK cell development in hIL-7 and hIL-15 knockin NOD\/SCID\/IL2rgKO mice","volume":"2","author":"Matsuda","year":"2019","journal-title":"Life Sci alliance"},{"key":"B107","doi-asserted-by":"publisher","first-page":"17230","DOI":"10.1038\/s41598-017-17442-7","article-title":"Long-term maintenance of peripheral blood derived human NK cells in a novel human IL-15- transgenic NOG mouse","volume":"7","author":"Katano","year":"2017","journal-title":"Sci Rep"},{"key":"B108","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1182\/blood-2020-141606","article-title":"Initial clinical activity of FT596, a first-in-class, multi-antigen targeted, off-the-shelf, iPSC-derived CD19 CAR NK cell therapy in relapsed\/refractory B-cell lymphoma","volume":"136","author":"Bachanova","year":"2020","journal-title":"Blood"},{"key":"B109","doi-asserted-by":"publisher","DOI":"10.1016\/S2352-3026(25)00142-5","article-title":"Off-the-shelf induced pluripotent stem-cell-derived natural killer-cell therapy in relapsed or refractory B-cell lymphoma: a multicentre, open-label, phase 1 study","volume":"12","author":"Strati","year":"2025","journal-title":"Lancet Haematol"},{"key":"B110","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.2000164","article-title":"LILRB1 intron 1 has a polymorphic regulatory region that enhances transcription in NK cells and recruits YY1","volume":"204","author":"Yu","year":"2020","journal-title":"J Immunol"},{"key":"B111","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2021.10.008","article-title":"Multiplex CRISPR\/Cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations","volume":"23","author":"Samuelson","year":"2021","journal-title":"Mol Ther Methods Clin Dev"},{"key":"B112","doi-asserted-by":"publisher","first-page":"4063","DOI":"10.1038\/s41467-019-11955-7","article-title":"Mitigation of off-target toxicity in CRISPR-Cas9 screens for essential non-coding elements","volume":"10","author":"Tycko","year":"2019","journal-title":"Nat Commun"},{"key":"B113","doi-asserted-by":"publisher","first-page":"7208","DOI":"10.1038\/s41467-025-62606-z","article-title":"The hidden risks of CRISPR\/Cas: structural variations and genome integrity","volume":"16","author":"Aussel","year":"2025","journal-title":"Nat Commun"},{"key":"B114","doi-asserted-by":"publisher","first-page":"212","DOI":"10.1038\/s41467-023-35886-6","article-title":"Assessing and advancing the safety of CRISPR-Cas tools: from DNA to RNA editing","volume":"14","author":"Tao","year":"2023","journal-title":"Nat Commun"},{"key":"B115","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-020-0453-z","article-title":"Phage-assisted evolution of an adenine base editor with improved Cas domain compatibility and activity","volume":"38","author":"Richter","year":"2020","journal-title":"Nat Biotechnol"},{"key":"B116","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2023.1339189","article-title":"Beyond the promise: evaluating and mitigating off-target effects in CRISPR gene editing for safer therapeutics","volume":"11","author":"Lopes","year":"2023","journal-title":"Front Bioeng Biotechnol"},{"key":"B117","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3852","article-title":"Genome-wide target specificities of CRISPR RNA-guided programmable deaminases","volume":"35","author":"Kim","year":"2017","journal-title":"Nat Biotechnol"},{"key":"B118","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-019-0050-1","article-title":"Genome-wide target specificity of CRISPR RNA-guided adenine base editors","volume":"37","author":"Kim","year":"2019","journal-title":"Nat Biotechnol"},{"key":"B119","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1038\/s41467-018-07988-z","article-title":"Genome-wide profiling of adenine base editor specificity by EndoV-seq","volume":"10","author":"Liang","year":"2019","journal-title":"Nat Commun"},{"key":"B120","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-1161-z","article-title":"Transcriptome-wide off-target RNA editing induced by CRISPR-guided DNA base editors","volume":"569","author":"Gr\u00fcnewald","year":"2019","journal-title":"Nature"},{"key":"B121","doi-asserted-by":"publisher","first-page":"882","DOI":"10.1038\/s42003-025-08275-6","article-title":"A versatile CRISPR\/Cas9 system off-target prediction tool using language model","volume":"8","author":"Du","year":"2025","journal-title":"Commun Biol"},{"key":"B122","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41551-025-01463-z","article-title":"CRISPR-GPT for agentic automation of gene-editing experiments","author":"Qu","year":"2025","journal-title":"Nat Biomed Eng"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1711414\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T06:28:55Z","timestamp":1764916135000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1711414\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,5]]},"references-count":122,"alternative-id":["10.3389\/fimmu.2025.1711414"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2025.1711414","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,5]]},"article-number":"1711414"}}