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By longitudinal monitoring of T cell functionality and dynamics in 15 healthy donors, we isolated 19 TCRs specific for Wilms\u2019 tumor antigen 1 (WT1), which is overexpressed by several tumor types. TCRs recognized several peptides restricted by common human leukocyte antigen (HLA) alleles and displayed a wide range of functional avidities. We selected five high-avidity HLA-A*02:01\u2013restricted TCRs, three that were specific to the less explored immunodominant WT1\n            <jats:sub>37\u201345<\/jats:sub>\n            and two that were specific to the noncanonical WT1\n            <jats:sub>\u221278\u201364<\/jats:sub>\n            epitopes, both naturally processed by primary acute myeloid leukemia (AML) blasts. With CRISPR-Cas9 genome editing tools, we combined TCR-targeted integration into the TCR \u03b1 constant (\n            <jats:italic>TRAC<\/jats:italic>\n            ) locus with TCR \u03b2 constant (\n            <jats:italic>TRBC<\/jats:italic>\n            ) knockout, thus avoiding TCR\u03b1\u03b2 mispairing and maximizing TCR expression and function. The engineered lymphocytes were enriched in memory stem T cells. A unique WT1\n            <jats:sub>37\u201345<\/jats:sub>\n            -specific TCR showed antigen-specific responses and efficiently killed AML blasts, acute lymphoblastic leukemia blasts, and glioblastoma cells in vitro and in vivo in the absence of off-tumor toxicity. T cells engineered to express this receptor are being advanced into clinical development for AML immunotherapy and represent a candidate therapy for other WT1-expressing tumors.\n          <\/jats:p>","DOI":"10.1126\/scitranslmed.abg8027","type":"journal-article","created":{"date-parts":[[2022,2,9]],"date-time":"2022-02-09T18:55:26Z","timestamp":1644432926000},"source":"Crossref","is-referenced-by-count":56,"title":["CRISPR-based gene disruption and integration of high-avidity, WT1-specific T cell receptors improve antitumor T cell function"],"prefix":"10.1126","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2954-8278","authenticated-orcid":true,"given":"Eliana","family":"Ruggiero","sequence":"first","affiliation":[{"name":"Experimental Hematology Unit, Division of Immunology, Transplantation and Infectious Diseases, Ospedale San Raffaele Scientific Institute, 20132 Milan, 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