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Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection
Open accessAbstract
Chimeric antigen receptors (CARs) are synthetic receptors that redirect and reprogram T cells to mediate tumour rejection. The most successful CARs used to date are those targeting CD19 (ref. 2), which offer the prospect of complete remission in patients with chemorefractory or relapsed B-cell malignancies. CARs are typically transduced into the T cells of a patient using γ-retroviral vectors or other randomly integrating vectors, which may result in clonal expansion, oncogenic transformation, variegated transgene expression and transcriptional silencing. Recent advances in genome editing enable efficient sequence-specific interventions in human cells, including targeted gene delivery to the CCR5 and AAVS1 loci. Here we show that directing a CD19-specific CAR to the T-cell receptor α constant (TRAC) locus not only results in uniform CAR expression in human peripheral blood T cells, but also enhances T-cell potency, with edited cells vastly outperforming conventionally generated CAR T cells in a mouse model of acute lymphoblastic leukaemia. We further demonstrate that targeting the CAR to the TRAC locus averts tonic CAR signalling and establishes effective internalization and re-expression of the CAR following single or repeated exposure to antigen, delaying effector T-cell differentiation and exhaustion. These findings uncover facets of CAR immunobiology and underscore the potential of CRISPR/Cas9 genome editing to advance immunotherapies.
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“AAV6 homologous donor DNA templates are an efficient way to deliver donor templates for sensitive primary immune cells, including HSPCs and T-Cells15,35–40.”
From A high-fidelity Cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human hematopoietic stem and progenitor cells (2018) · cites this paper as [39]
Experimental — accuracy not yet measuredIn the citing paper
“More recent work has revealed potential advantages of targeted integrations of antigen receptor genes into defined genomic loci (Eyquem et al., 2017), which can be achieved with programmable nucleases such as CRISPR-Cas9 along with viral or non-viral knock-in templates (Gwiazda et al., 2016; Eyquem et al., 2017; Roth et al., 2018).”
From Pooled Knockin Targeting for Genome Engineering of Cellular Immunotherapies (2020) · cites this paper as Eyquem et al., 2017
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Citations
Cites
Molecular and cellular insights into T cell exhaustion
Wherry, E. J., & Kurachi, M. (2015) · Nature Reviews Immunology · doi:10.1038/nri3862 · Cited by 4,652
Brentjens, R. J., Davila, M. L., Riviere, I., Park, J., Wang, X., Cowell, L. G., Bartido, S., Stefanski, J., Taylor, C., Olszewska, M., et al. (2013) · Science Translational Medicine · doi:10.1126/scitranslmed.3005930 · Cited by 2,063
Self-citationBlackburn, S. D., Shin, H., Haining, W. N., Zou, T., Workman, C. J., Polley, A., Betts, M. R., Freeman, G. J., Vignali, D. A. A., & Wherry, E. J. (2009) · Nature Immunology · doi:10.1038/ni.1679 · Cited by 2,062
A human memory T cell subset with stem cell–like properties
Gattinoni, L., Lugli, E., Ji, Y., Pos, Z., Paulos, C. M., Quigley, M. F., Almeida, J. R., Gostick, E., Yu, Z., Carpenito, C., et al. (2011) · Nature Medicine · doi:10.1038/nm.2446 · Cited by 1,974
Long, A. H., Haso, W. M., Shern, J. F., Wanhainen, K. M., Murgai, M., Ingaramo, M., Smith, J. P., Walker, A. J., Kohler, M. E., Venkateshwara, V. R., et al. (2015) · Nature Medicine · doi:10.1038/nm.3838 · Cited by 1,815
Human T-lymphocyte cytotoxicity and proliferation directed by a single chimeric TCRζ /CD28 receptor
Maher, J. F., Brentjens, R. J., Gunset, G., Rivière, I., & Sadelain, M. (2002) · Nature Biotechnology · doi:10.1038/nbt0102-70 · Cited by 1,031
Self-citationBiology and Applications of CRISPR Systems: Harnessing Nature’s Toolbox for Genome Engineering
Wright, A. V., Nuñez, J. K., & Doudna, J. A. (2016) · Cell · doi:10.1016/j.cell.2015.12.035 · Cited by 989
Sommermeyer, D., Hudecek, M., Kosasih, P. L., Gogishvili, T., Maloney, D. G., Turtle, C. J., & Riddell, S. R. (2015) · Leukemia · doi:10.1038/leu.2015.247 · Cited by 890
Zhao, Z., Condomines, M., van der Stegen, S. J. C., Perna, F., Kloss, C. C., Gunset, G., Plotkin, J., & Sadelain, M. (2015) · Cancer Cell · doi:10.1016/j.ccell.2015.09.004 · Cited by 843
Self-citationTolerance and exhaustion: defining mechanisms of T cell dysfunction
Schietinger, A., & Greenberg, P. D. (2013) · Trends in Immunology · doi:10.1016/j.it.2013.10.001 · Cited by 704
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Items 501–600 of 1,002
CRISPR/Cas9 in Cancer Immunotherapy: Animal Models and Human Clinical Trials
Khalaf, K., Janowicz, K., Dyszkiewicz-Konwińska, M., Hutchings, G., Dompé, C., Moncrieff, L., Jankowski, M., Machnik, M., Oleksiewicz, U., Kocherova, I., et al. (2020) · Genes · doi:10.3390/genes11080921 · Cited by 52
“In vivo comparison at TRAC locus in an Acute Lymphoblastic Leukemia (ALL) mouse model performed between cells with retroviral vector chimeric antigen receptors (RV-CARs), TRAC-CARs cells, and TCR-disrupted cells expressing chimeric antigen receptor [(RV-CAR(+)-TCR(-))], demonstrated greater response and prolonged surv…”
BackgroundStem cell-like memory T cells: The generation and application
Wang, Y., Qiu, F., Xu, Y., Hou, X., Zhang, Z., Huang, L., Wang, H., Xing, H., & Wu, S. (2021) · Journal of Leukocyte Biology · doi:10.1002/jlb.5mr0321-145r · Cited by 49
Michaels, Y. S., Edgar, J. M., Major, M. C., Castle, E. L., Zimmerman, C., Yin, T., Hagner, A., Lau, C., Hsu, H. H., Ibanez-Rios, M. I., et al. (2022) · Science Advances · doi:10.1126/sciadv.abn5522 · Cited by 53
García-Calderón, C. B., Sierro-Martínez, B., García‐Guerrero, E., Sanoja-Flores, L., Muñoz-García, R., Ruiz-Maldonado, V., Jiménez‐León, M. R., Delgado‐Serrano, J., Molinos-Quintana, Á., Guijarro-Albaladejo, B., et al. (2023) · Frontiers in Immunology · doi:10.3389/fimmu.2023.1152498 · Cited by 51
“Several studies have analyzed exhaustion markers in CAR-T cells at the time of leukapheresis and in the infusion products demonstrating that increased exhaustion markers at these time points correlate with reduce efficacy of CAR-T cells (6, 9, 12, 13, 33, 52, 53).”
BackgroundThe transformative potential of AI-driven CRISPR-Cas9 genome editing to enhance CAR T-cell therapy
Boretti, A. A. (2024) · Computers in Biology and Medicine · doi:10.1016/j.compbiomed.2024.109137 · Cited by 53
Cost-effective strategies for CAR-T cell therapy manufacturing
Abdo, L., Batista-Silva, L. R., & Bonamino, M. H. (2025) · Molecular Therapy Oncology · doi:10.1016/j.omton.2025.200980 · Cited by 54
“37 Additionally, this approach may help mitigate any unintended effects associated with nonspecific recognition through the T cell receptor (TCR).”
CRISPR therapeutic tools for complex genetic disorders and cancer (Review)
Baliou, S., Adamaki, M., Kyriakopoulos, A. M., Spandidos, D. A., Panayiotidis, M. I., Christodoulou, I., & Zoumpourlis, V. C. (2018) · International Journal of Oncology · doi:10.3892/ijo.2018.4434 · Cited by 55
“For example, Eyquem et al demonstrated that Cas9-modified effector T cells displayed CD19-specific CAR receptor, recognizing the specific sequence of T cell receptor alpha constant (TRAC) and successfully treating acute lymphoblastic leukemia in a mouse model (160).”
Genomics-Guided Immunotherapy for Precision Medicine in Cancer
Mukherjee, S. (2019) · Cancer Biotherapy and Radiopharmaceuticals · doi:10.1089/cbr.2018.2758 · Cited by 50
“In a mouse model of lymphoblastic leukemia, CRISPR/Cas9-edited CAR T cells were shown to reduce T cell exhaustion relative to conventional CAR T cells.(94) However, due to potential off-target effects of CRISPR/Cas9, which may change based on the context of genetic background, it has been challenging to apply CRISPR/…”
BackgroundGene Editing of Hematopoietic Stem Cells: Hopes and Hurdles Toward Clinical Translation
Ferrari, S., Vavassori, V., Canarutto, D., Jacob, A., Castiello, M. C., Omer, A., & Genovese, P. (2021) · Frontiers in Genome Editing · doi:10.3389/fgeed.2021.618378 · Cited by 52
“, 2016) have become the gold standard to achieve a high but transient nuclease activity in HSPCs and other target cells (Hubbard et al., 2016; Eyquem et al., 2017).”
BackgroundAllogeneic gamma delta T cells as adoptive cellular therapy for hematologic malignancies
Jhita, N., & Raikar, S. S. (2022) · Exploration of Immunology · doi:10.37349/ei.2022.00054 · Cited by 51
“The different gene editing tools that have used in this setting include Zinc finger nucleases (ZFN) [76–78], transcription activator-like effector nucleases (TALEN) [79–81], and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 genome editing [82, 83].”
CAR-engineered lymphocyte persistence is governed by a FAS ligand–FAS autoregulatory circuit
Yi, F., Cohen, T., Zimmerman, N., Dündar, F., Zumbo, P., Eltilib, R., Brophy, E. J., Arkin, H., Feucht, J., Gormally, M. V., et al. (2025) · Nature Cancer · doi:10.1038/s43018-025-01009-x · Cited by 55
Self-citationVo, M.-C., Tran, V.-D.-H., Nguyen, V., Ruzimurodov, N., Trung, D. T., Kim, S. K., Jung, S., & Lee, J. (2025) · Journal of Hematology & Oncology · doi:10.1186/s13045-025-01744-9 · Cited by 59
“In addition, CRISPR-Cas9 gene editing has been used to delete inhibitory regulators, such as TET2, RASA2, and REGNASE-1, or to overexpress transcription factors like c-Jun, improving CAR-T cell persistence and effector function [240].”
Evolution of chimeric antigen receptor (CAR) T cell therapy: current status and future perspectives
Lee, Y.-H., & Kim, C. H. (2019) · Archives of Pharmacal Research · doi:10.1007/s12272-019-01136-x · Cited by 50
Marofi, F., Rahman, H. S., Achmad, M. H., Sergeevna, K. N., Suksatan, W., Abdelbasset, W. K., Mikhailova, M. V., Shomali, N., Yazdanifar, M., Hassanzadeh, A., et al. (2021) · Frontiers in Immunology · doi:10.3389/fimmu.2021.681984 · Cited by 48
RetractedEngineering strategies for broad application of TCR-T- and CAR-T-cell therapies
Watanabe, K., & Nishikawa, H. (2021) · International Immunology · doi:10.1093/intimm/dxab052 · Cited by 50
RETRACTED ARTICLE: Hurdles to breakthrough in CAR T cell therapy of solid tumors
Marofi, F., Achmad, H., Bokov, D., Abdelbasset, W. K., Alsadoon, Z., Chupradit, S., Suksatan, W., Shariatzadeh, S., Hasanpoor, Z., Yazdanifar, M., et al. (2022) · Stem Cell Research & Therapy · doi:10.1186/s13287-022-02819-x · Cited by 47
“I C Production of anti-human PD1-blocking scFv by CD19 or MUC16 ecto -modified CAR T cells augmented their killing activity against PDL1 + tumor cells [123].”
InfluentialRetractedJanelle, V., & Delisle, J. (2021) · Cancers · doi:10.3390/cancers13040598 · Cited by 52
Sun, T., Li, Y., Yang, Y., Liu, B., Cao, Y., & Yang, W. (2022) · Cell Death and Disease · doi:10.1038/s41419-022-05319-1 · Cited by 47
“directed a CAR to the T-cell receptor α constant (TRAC) locus to induce uniform CAR expression in human peripheral blood T cells and enhance T-cell potency [33].”
BackgroundAnthony-Gonda, K., Ray, A. K., Su, H., Wang, Y., Xiong, Y., Lee, D. M., Block, A., Chilunda, V., Weiselberg, J., Zemelko, L., et al. (2022) · JCI Insight · doi:10.1172/jci.insight.161698 · Cited by 47
“with reports that promoter identity may augment the therapeutic efficacy of the CAR T cell product (37),”
Antibody with Infinite Affinity for In Vivo Tracking of Genetically Engineered Lymphocytes
Krebs, S. S., Ahad, A., Carter, L. M., Eyquem, J., Brand, C., Bell, M. M., Ponomarev, V., Reiner, T., Meares, C. F., Gottschalk, S. M., et al. (2018) · Journal of Nuclear Medicine · doi:10.2967/jnumed.118.208041 · Cited by 47
“Cytotoxicity Assays The cytotoxicity of T cells transduced with CD19-CAR was determined by standard luciferase-based assay using NALM-6–expressing firefly luciferase–GFP as target cells (24).”
MethodSelf-citationCRISPR/Cas9 System: A Bacterial Tailor for Genomic Engineering
Lone, B. A., Karna, S. K. L., Ahmad, F., Shahi, N., & Pokharel, Y. R. (2018) · Genetics Research International · doi:10.1155/2018/3797214 · Cited by 48
“This technology is also applied for CD19-specific CAR coding sequence into the T cell receptor α constant (TRAC) locus to create CAR T cells for cancer therapy [131].”
MethodWendel, P., Reindl, L. M., Bexte, T., Künnemeyer, L., Särchen, V., Albinger, N., Mackensen, A., Rettinger, E., Bopp, T., & Ullrich, E. (2021) · Cancers · doi:10.3390/cancers13061481 · Cited by 48
“Furthermore, CRISPR/Cas9 can be used to direct the CAR construct to a known and non-random genomic location to disrupt the endogenous TCR locus [110].”
BackgroundLi, H., Zhou, X., Wang, G., Hua, D., Li, S., Xu, T., Dong, M., Cui, X., Yang, X., Wu, Y., et al. (2021) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/clinem/dgab819 · Cited by 50
Jiang, L., Ingelshed, K., Shen, Y., Boddul, S. V., Iyer, V. S., Kasza, Z., Sedimbi, S. K., Lane, D. P., & Wermeling, F. (2021) · Cancer Research · doi:10.1158/0008-5472.can-21-1692 · Cited by 47
“CRISPR-mediated modifications of cells used for chimeric antigen receptor (CAR)-based immunotherapy is another clinical setting where CRISPR is likely to have a large impact (12).”
BackgroundUnleashing the Therapeutic Potential of CAR-T Cell Therapy Using Gene-Editing Technologies.
Jung, I.-Y., & Lee, J. (2018) · Europe PMC (PubMed Central) · doi:10.14348/molcells.2018.0242 · Cited by 49
“However, strong, unregulated CAR expression can result in accelerated T-cell differentiation and exhaustion (Eyquem et al., 2017).”
BackgroundMethodCancer gene therapy goes viral: viral vector platforms come of age
Bezeljak, U. (2022) · Radiology and Oncology · doi:10.2478/raon-2022-0002 · Cited by 48
Concise Review: Human Pluripotent Stem Cells to Produce Cell-Based Cancer Immunotherapy
Huang, Z., Lai, Y.-S., Li, Y., Blum, R. H., & Kaufman, D. S. (2017) · Stem Cells · doi:10.1002/stem.2754 · Cited by 48
T cell engineering for adoptive T cell therapy: safety and receptor avidity
D’Ippolito, E., Schober, K., Nauerth, M., & Busch, D. H. (2019) · Cancer Immunology Immunotherapy · doi:10.1007/s00262-019-02395-9 · Cited by 47
Ansell, S. M., & Lin, Y. (2020) · Journal of Clinical Investigation · doi:10.1172/jci129206 · Cited by 48
Okamura, T., Yamamoto, K., & Fujio, K. (2018) · Frontiers in Immunology · doi:10.3389/fimmu.2018.00340 · Cited by 41
“Thus, it increased the therapeutic potency of engineered T cells (106).”
BackgroundBalke-Want, H., Keerthi, V., Gkitsas, N., Mancini, A., Kurgan, G., Fowler, C., Xu, P., Liu, X.-K., Asano, K., Patel, S., et al. (2023) · Molecular Cancer · doi:10.1186/s12943-023-01799-7 · Cited by 41
“CRISPR/Cas9 with adeno-associated virus 6 (AAV6) can deliver sitedirected modifications of the genome via HDR, and this approach may reduce the risk of insertional mutagenesis compared to random insertions delivered via retroviruses and simultaneously knockout a gene of interest [18, 19].”
BackgroundChimeric Antigen Receptor T-Cells for the Treatment of B-Cell Acute Lymphoblastic Leukemia
Tomuleasa, C., Fuji, S., Berce, C., Onaciu, A., Chira, S., Petrushev, B., Micu, W.-T., Moisoiu, V., Osan, C., Constantinescu, C. S., et al. (2018) · Frontiers in Immunology · doi:10.3389/fimmu.2018.00239 · Cited by 46
“Up to date, CRISPR/Cas9 already proved its applicability in the field of immunotherapy by enhancing CAR T-cells potency by knockout diverse genes to improve target recognition and cytotoxic activity (104).”
InfluentialMechanisms of and approaches to overcoming resistance to immunotherapy
Schultz, L. M., & Gardner, R. A. (2019) · Hematology · doi:10.1182/hematology.2019000018 · Cited by 45
“Furthermore, TRACCAR T cells are protected from accelerated exhaustion and yield improved antitumor activity in animal models.(43)”
BackgroundCRISPR Gene Editing of Human Primary NK and T Cells for Cancer Immunotherapy
Elmas, E., Saljoughian, N., Pereira, M. D. S. F., Tullius, B. P., Sorathia, K., Nakkula, R. J., Lee, D. A., & Kararoudi, M. N. (2022) · Frontiers in Oncology · doi:10.3389/fonc.2022.834002 · Cited by 47
“Next, the HDR template encoding CD19 CAR was delivered to the cells via AAV6 transduction (60).”
MethodSite-specific transgene integration in chimeric antigen receptor (CAR) T cell therapies
Dabiri, H., Kozani, P. S., Habibi‐Anbouhi, M., Godarzee, M. M., Haddadi, M. H., Basiri, M., Ziaei, V., Sadeghizadeh, M., & Saffar, E. H. (2023) · Biomarker Research · doi:10.1186/s40364-023-00509-1 · Cited by 45
“Therefore, the TCR locus can be considered one of the most interesting gene targets for the integration of the CAR transgene [25].”
BackgroundMethodInfluentialAllemailem, K. S., Alsahli, M. A., Almatroudi, A., Alrumaihi, F., Abdulmonem, W. A., Moawad, A. A., Alwanian, W. M., Almansour, N. M., Rahmani, A. H., & Khan, A. A. (2023) · International Journal of Nanomedicine · doi:10.2147/ijn.s424872 · Cited by 49
Synthetic manipulation of the cancer-immunity cycle: CAR-T cell therapy
Singh, N., & Maus, M. V. (2023) · Immunity · doi:10.1016/j.immuni.2023.09.010 · Cited by 44
Genetic ablation of adhesion ligands mitigates rejection of allogeneic cellular immunotherapies
Hammer, Q., Perica, K., Mbofung, R. M., van Ooijen, H., Martin, K. E., Momayyezi, P., Varady, E., Pan, Y., Jelcic, M., Groff, B. C., et al. (2024) · Cell stem cell · doi:10.1016/j.stem.2024.06.011 · Cited by 54
Self-citationAllogeneic “Off-the-Shelf” CAR T cells: Challenges and advances
Chen, S., & van den Brink, M. R. M. (2024) · Best Practice & Research Clinical Haematology · doi:10.1016/j.beha.2024.101566 · Cited by 51
In vivo site-specific engineering to reprogram T cells
Nyberg, W. A., Bernard, P.-L., Ngo, W., Wang, C. H., Ark, J., Rothrock, A., Borgo, G. M., Kimmerly, G. R., Jung, J. H., Allain, V., et al. (2026) · Nature · doi:10.1038/s41586-026-10235-x · Cited by 55
Self-citationAdoptive T Cell Therapy: New Avenues Leading to Safe Targets and Powerful Allies
Hammerl, D., Rieder, D., Martens, J. W. M., Trajanoski, Z., & Debets, R. (2018) · Trends in Immunology · doi:10.1016/j.it.2018.09.004 · Cited by 46
Bertaina, A., & Roncarolo, M. G. (2019) · Frontiers in Immunology · doi:10.3389/fimmu.2019.01342 · Cited by 45
“Furthermore, targeted integration of the CAR into the constant region of the TRAC in-frame with the endogenous gene’s open reading frame has the potential to increase CAR T cell functionality and persistence (97).”
BackgroundAllen, M. E., Zhou, W., Thangaraj, J., Kyriakakis, P., Wu, Y., Huang, Z., Ho, P., Pan, Y., Limsakul, P., Xu, X., et al. (2019) · ACS Synthetic Biology · doi:10.1021/acssynbio.9b00175 · Cited by 43
Expanding CAR T cells in human platelet lysate renders T cells with in vivo longevity
Chavez, A. G. T., McKenna, M. K., Canestrari, E., Dann, C. T., Ramos, C. A., Lulla, P., Leen, A. M., Vera, J. F., & Watanabe, N. (2019) · Journal for ImmunoTherapy of Cancer · doi:10.1186/s40425-019-0804-9 · Cited by 45
Kawai, Y., Kawana‐Tachikawa, A., Kitayama, S., Ueda, T., Miki, S., Watanabe, A., & Kaneko, S. (2021) · Molecular Therapy · doi:10.1016/j.ymthe.2021.05.016 · Cited by 43
CRISPR-Cas System: The Current and Emerging Translational Landscape
Bhokisham, N., Laudermilch, E., Traeger, L. L., Bonilla, T. D., Ruíz-Estévez, M., & Becker, J. R. (2023) · Cells · doi:10.3390/cells12081103 · Cited by 46
“In addition, an CD19-specific CAR cassette was precisely inserted at the TRAC loci using the CRISPR-Cas9 system to regulate the expression of CAR [75].”
MethodInfluentialAltered cancer metabolism and implications for next-generation CAR T-cell therapies
Ramapriyan, R., Vykunta, V. S., Vandecandelaere, G., Richardson, L. G., Sun, J. M., Curry, W. T., & Choi, B. D. (2024) · Pharmacology & Therapeutics · doi:10.1016/j.pharmthera.2024.108667 · Cited by 48
Clauss, J., Obenaus, M., Miskey, C., Ivics, Z., Izsvák, Z., Uckert, W., & Bunse, M. (2018) · MDC Repository (Max-Delbrueck-Center for Molecular Medicine) · doi:10.1089/hum.2017.136 · Cited by 1
“49 However, in case of TCR-engineered T cells, effective inhibition of mixed TCR dimer formation requires not only the knockdown of both endogenous TCR chains but also a laborious strategy to identify and isolate cells with the desired genotype.”
From synthetic biology to human therapy: engineered mammalian cells
Scheller, L., & Fussenegger, M. (2019) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2019.02.023 · Cited by 43
Using Gene Editing Approaches to Fine-Tune the Immune System
Pavlovic, K., Tristán‐Manzano, M., Maldonado‐Pérez, N., Cortijo-Gutierréz, M., Sánchez-Hernández, S., Justicia-Lirio, P., Carmona-Luque, M. D., Herrera, C., Martín, F., & Benabdellah, K. (2020) · Frontiers in Immunology · doi:10.3389/fimmu.2020.570672 · Cited by 49
“developed a similar strategy to incorporate the CAR construct into the TRAC locus (76).”
BackgroundMethodT-cell–engaging Therapy for Solid Tumors
de Miguel, M., Umaña, P., de Morais, A. L. G., Moreno, V., & Calvo, E. (2020) · Clinical Cancer Research · doi:10.1158/1078-0432.ccr-20-2448 · Cited by 42
“A new approach is using electroporation together with CRISPR/Cas9 for replacement of the endogenous TCR by the recombinant construct, which is being developed successfully (85).”
MethodUniversal toxin-based selection for precise genome engineering in human cells
Li, S., Akrap, N., Cerboni, S., Porritt, M. J., Wimberger, S., Lundin, A., Möller, C., Firth, M. A., Gordon, E., Lazovic, B., et al. (2021) · Nature Communications · doi:10.1038/s41467-020-20810-z · Cited by 46
New cell sources for CAR-based immunotherapy
Mazinani, M., & Rahbarizadeh, F. (2023) · Biomarker Research · doi:10.1186/s40364-023-00482-9 · Cited by 43
“This dual strategy not only averts tonic CAR signaling by optimizing CAR internalization and re-expression kinetics but also minimizes the risk of GvHD by diminishing the expression of αβTCRs on the T cell surface [25].”
BackgroundLipid nanoparticle-based mRNA vaccines: a new frontier in precision oncology
Jacob, E. M., Huang, J., & Chen, M. (2024) · Precision Clinical Medicine · doi:10.1093/pcmedi/pbae017 · Cited by 48
“As an alternative to integrating viral vectors, elec-tr opor ation ( EP ) of CAS9 mRNA into human T cells allows directed integration of a CD19-specific CAR to the T cell receptor α constant ( TRAC ) locus, resulting in constant CAR expression as well as im-pr ov ed T cell potency [ 70 ].”
MethodYe, B., Smerin, D., Qingping, G., Kang, C., & Xiong, X. (2017) · Cancer Letters · doi:10.1016/j.canlet.2017.12.017 · Cited by 41
Heyman, B. M., & Yang, Y. (2019) · Cancers · doi:10.3390/cancers11020191 · Cited by 43
“Mechanistically, the investigators found that specific targeting to the TRAC locus prevents tonic CAR signaling, improving persistence, delaying effector T-cell differentiation and exhaustion [130].”
BackgroundMethodMechanisms of failure of chimeric antigen receptor T-cell therapy
Li, X., & Chen, W. (2019) · Current Opinion in Hematology · doi:10.1097/moh.0000000000000548 · Cited by 41
Nanotechnology Promotes Genetic and Functional Modifications of Therapeutic T Cells Against Cancer
Abdalla, A. M. E., Xiao, L. J., Miao, Y., Huang, L., Fadlallah, G. M., Gauthier, M., Ouyang, C., & Yang, G. (2020) · Advanced Science · doi:10.1002/advs.201903164 · Cited by 42
Thangam, R., Patel, K. D., Kang, H., & Paulmurugan, R. (2021) · Vaccines · doi:10.3390/vaccines9080935 · Cited by 44
“, IFN-γ) are predominant molecules that link the innate and adaptive immunity in patients for the uptake and exposure of naive T-cells in the activation of tumor-specific CTL responses [34,35].”
BackgroundEngineering stem cells for cancer immunotherapy
Li, Y.-R., Zhou, Y., Kramer, A. B., & Yang, L. (2021) · Trends in cancer · doi:10.1016/j.trecan.2021.08.004 · Cited by 42
“Insertion of the CAR gene construct into the TRAC locus has been accomplished by inserting the CAR construct and knocking out the TCR gene in one step [39].”
MethodRNA Therapeutics for Improving CAR T-cell Safety and Efficacy
Schaible, P., Bethge, W. A., Lengerke, C., & Haraszti, R. Á. (2022) · Cancer Research · doi:10.1158/0008-5472.can-22-2155 · Cited by 43
Engineered Treg cells: The heir to the throne of immunotherapy
Stucchi, A., Maspes, F., Montee-Rodrigues, E., & Fousteri, G. (2023) · Journal of Autoimmunity · doi:10.1016/j.jaut.2022.102986 · Cited by 43
Enhancing cellular immunotherapies in cancer by engineering selective therapeutic resistance
Wellhausen, N., Baek, J. S., Gill, S. I., & June, C. H. (2024) · Nature reviews. Cancer · doi:10.1038/s41568-024-00723-5 · Cited by 45
“In contrast, CRISPR– Cas9 knock-in or prime editing-assisted gene insertion allow for the placement of transgenes under endogenous transcriptional control 132,133 .”
Xia, Z., Jin, Q., Long, Z., He, Y., Liu, F., Sun, C., Liao, J., Wang, C., Wang, C., Zheng, J., et al. (2024) · Nature Biomedical Engineering · doi:10.1038/s41551-024-01258-8 · Cited by 43
Toward the development of true “off‐the‐shelf” synthetic T‐cell immunotherapy
Iriguchi, S., & Kaneko, S. (2018) · Cancer Science · doi:10.1111/cas.13892 · Cited by 43
Chimeric Antigen Receptor Cell Therapy: Overcoming Obstacles to Battle Cancer
Petty, A. J., Heyman, B. M., & Yang, Y. (2020) · Cancers · doi:10.3390/cancers12040842 · Cited by 40
“were able to direct a CD19-targeting CAR to the T cell receptor constant (TRAC) locus, resulting in enhanced potency and antitumor activity [74].”
BackgroundNew directions in chimeric antigen receptor T cell [CAR‐T] therapy and related flow cytometry
Maryamchik, E., Gallagher, K., Preffer, F. I., Kadauke, S., & Maus, M. V. (2020) · Cytometry Part B Clinical Cytometry · doi:10.1002/cyto.b.21880 · Cited by 43
“enables the cells to regulate CAR expression in response to the antigen present, thus delaying terminal T-cell differentiation and exhaustion (Eyquem et al., 2017; MacLeod et al., 2017).”
BackgroundCRISPR screen in mechanism and target discovery for cancer immunotherapy
Liu, D., Zhao, X., Tang, A., Xu, X., Liu, S., Zha, L., Ma, W., Zheng, J., & Shi, M. (2020) · Biochimica et Biophysica Acta (BBA) - Reviews on Cancer · doi:10.1016/j.bbcan.2020.188378 · Cited by 44
CRISPR-based strategies for targeted transgene knock-in and gene correction
Lau, C., Tin, C., & Suh, Y. (2020) · Faculty Reviews · doi:10.12703/r/9-20 · Cited by 46
Regulatory T Cell Therapy of Graft-versus-Host Disease: Advances and Challenges
Hefazi, M., Bolivar-Wagers, S., & Blazar, B. R. (2021) · International Journal of Molecular Sciences · doi:10.3390/ijms22189676 · Cited by 40
Branella, G. M., & Spencer, H. T. (2021) · Cells · doi:10.3390/cells11010021 · Cited by 42
“Many scenarios can lead to CAR tonic signaling besides scFv aggregation, including the level of CAR expression, certain costimulatory domains [24], and the addition of endogenous T-cell receptor (TCR) signaling in the CAR T cell [41].”
BackgroundEpitope-engineered human hematopoietic stem cells are shielded from CD123-targeted immunotherapy
Marone, R., Landmann, E., Devaux, A., Lepore, R., Seyres, D., Zuin, J., Burgold, T., Engdahl, C., Capoferri, G., Dell’Aglio, A., et al. (2023) · The Journal of Experimental Medicine · doi:10.1084/jem.20231235 · Cited by 39
“Hence, various strategies to increase CAR T longevity are explored (Eyquem et al., 2017; Lynn et al., 2019).”
Progress and pitfalls of gene editing technology in CAR-T cell therapy: a state-of-the-art review
Moradi, V., Khodabandehloo, E., Alidadi, M., Omidkhoda, A., & Ahmadbeigi, N. (2024) · Frontiers in Oncology · doi:10.3389/fonc.2024.1388475 · Cited by 42
“Nevertheless, the risk of developing life-threatening GvHD and immunogenicity are the two main bottlenecks of allogeneic CAR-T cell therapy (94, 95).”
BackgroundInfluentialManipulating the tumor microenvironment by adoptive cell transfer of CAR T-cells
Gowrishankar, K., Birtwistle, L., & Micklethwaite, K. P. (2018) · Mammalian Genome · doi:10.1007/s00335-018-9756-5 · Cited by 43
Mirzaei, H. R., Jamali, A., Jafarzadeh, L., Masoumi, E., Alishah, K., Fallah-Mehrjardi, K., Emami, S. A. H., Noorbakhsh, F., Till, B. G., & Hadjati, J. (2018) · Journal of Cellular Physiology · doi:10.1002/jcp.27599 · Cited by 40
CAR T Cells: A Snapshot on the Growing Options to Design a CAR
Holzinger, A., & Abken, H. (2019) · HemaSphere · doi:10.1097/hs9.0000000000000172 · Cited by 40
CRISPR to fix bad blood: a new tool in basic and clinical hematology
González-Romero, E., Martínez-Valiente, C., García-Ruiz, C., Vázquez‐Manrique, R. P., Cervera, J., & Sanjuán-Pla, A. (2019) · Haematologica · doi:10.3324/haematol.2018.211359 · Cited by 42
Delivery of nucleic acid therapeutics for cancer immunotherapy
Zhou, S., Chen, W., Cole, J., & Zhu, G. (2020) · Medicine in Drug Discovery · doi:10.1016/j.medidd.2020.100023 · Cited by 42
“It has been demonstrated that directing a CD19specific CAR to the TRAC locus not only resulted in uniform CAR expression in human peripheral blood T cells but also enhanced T-cell potency; these edited cells largely outperformed conventionally generated CAR T cells in a mouse model of acute lymphoblastic leukemia [83].”
BackgroundExploiting the CRISPR‐Cas9 gene‐editing system for human cancers and immunotherapy
Afolabi, L. O., Afolabi, M. O., Sani, M. M., Okunowo, W. O., Yan, D., Chen, L., Zhang, Y., & Wan, X. (2021) · Clinical & Translational Immunology · doi:10.1002/cti2.1286 · Cited by 46
“The knock-in of CAR at the a constant locus of TCR improved T-cell antitumor activity.(64) Similarly, the integration of CAR into the TRAC locus prevented CAR signalling and immune cell”
BackgroundMethodOptimization of AAV6 transduction enhances site-specific genome editing of primary human lymphocytes
Rogers, G. L., Huang, C., Clark, R. D. E., Seclén, E., Chen, H.-Y., & Cannon, P. M. (2021) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2021.09.003 · Cited by 41
Development of CAR T Cell Therapy in Children—A Comprehensive Overview
Boettcher, M., Joechner, A., Li, Z., Yang, S. F., & Schlegel, P. (2022) · Journal of Clinical Medicine · doi:10.3390/jcm11082158 · Cited by 42
“Keywords: evolution of CAR T cells; FDA-approved CAR products; TcR versus CAR; limitations and complications of CAR T cell therapy; future directions of CAR T cell therapy”
BackgroundMethodResultInfluentialEngineering Next-Generation CAR-T Cells: Overcoming Tumor Hypoxia and Metabolism
Gao, T. A., & Chen, Y. Y. (2022) · Annual Review of Chemical and Biomolecular Engineering · doi:10.1146/annurev-chembioeng-092120-092914 · Cited by 41
Jo, D., Kaczmarek, S., Shin, O., Wang, L., Cowan, J., McComb, S., & Lee, S.-H. (2023) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2023.03.006 · Cited by 39
VanderBurgh, J. A., Corso, T. N., Levy, S. L., & Craighead, H. G. (2023) · Scientific Reports · doi:10.1038/s41598-023-33941-2 · Cited by 44
The Interplay between T Cells and Cancer: The Basis of Immunotherapy
Chen, C., Liu, X., Chang, C., Wang, H. Y., & Wang, R. (2023) · Genes · doi:10.3390/genes14051008 · Cited by 42
“By targeting the integration of the CAR construct into the TCR α constant region, universal expression of CAR and prolonged T cell persistence in vivo can be achieved [405,406].”
A Roadmap of CAR-T-Cell Therapy in Glioblastoma: Challenges and Future Perspectives
Montoya, M., Gallus, M., Phyu, S. M., Haegelin, J., de Groot, J., & Okada, H. (2024) · Cells · doi:10.3390/cells13090726 · Cited by 43
“To address this, groups have placed the CAR construct expression under the control of the T-cell receptor alpha constant (TRAC) locus, which knocks out the endogenous TCR, addressing the issue of GvHD [180,181].”
MethodThe role of allogeneic HSCT after CAR T cells for acute lymphoblastic leukemia
Jacoby, E. (2019) · Bone Marrow Transplantation · doi:10.1038/s41409-019-0604-3 · Cited by 39
Delivery methods for CRISPR/Cas9 gene editing in crustaceans
Xu, S., Pham, T. P., & Neupane, S. (2019) · Marine Life Science & Technology · doi:10.1007/s42995-019-00011-4 · Cited by 39
Padayachee, J., & Singh, M. (2020) · Nanobiomedicine · doi:10.1177/1849543520983196 · Cited by 39
“Introduction of the anti-CD19 CAR into the TCR locus has been shown using CRISPR, leading to effective CAR expression while avoiding T-cell exhaustion.(142) Multiplex strategies, allowing simultaneous knock out of TCRs, beta-2 microglobulin (B2 M), a subunit of HLA-I, and other proteins including PD-1 and CTLA-4, have…”
BackgroundReview of applications of CRISPR-Cas9 gene-editing technology in cancer research
Zhao, Z., Li, C., Tong, F., Deng, J., Huang, G., & Sang, Y. (2021) · Biological Procedures Online · doi:10.1186/s12575-021-00151-x · Cited by 42
“Furthermore, the edited cells performed much better than the conventionally generated CAR-T cells in the mouse model of acute lymphoblastic leukemia [88].”
BackgroundCRISPR/Cas9 as a therapeutic tool for triple negative breast cancer: from bench to clinics
Tiwari, P. K., Ko, T.-H., Dubey, R., Chouhan, M., Tsai, L.-W., Singh, H. N., Chaubey, K. K., Dayal, D., Chiang, C., & Kumar, S. (2023) · Frontiers in Molecular Biosciences · doi:10.3389/fmolb.2023.1214489 · Cited by 42
“Multiplex techniques have been developed, which allow the simultaneous elimination of TCRs, beta-2 microglobulin (B2 M), a subunit of HLA-I, and other proteins like PD-1 and CTLA-4, leading to produce allogeneic CAR T-cells, which may have higher anticancer property against TNBC (Eyquem et al., 2017; Liu et al., 2017;…”
MethodDevelopment of Beta-Amyloid-Specific CAR-Tregs for the Treatment of Alzheimer’s Disease
Saetzler, V., Riët, T., Schienke, A., Henschel, P., Freitag, K., Haake, A. F. H., Heppner, F. L., Buitrago‐Molina, L. E., Noyan, F., Jaeckel, E., et al. (2023) · Cells · doi:10.3390/cells12162115 · Cited by 39
“It has been demonstrated that this phenomenon is beneficial for the CAR function in T cells [47].”
BackgroundCRISPR/Cas9 system: recent applications in immuno-oncology and cancer immunotherapy
Chen, C., Wang, Z., & Qin, Y. (2023) · Experimental Hematology and Oncology · doi:10.1186/s40164-023-00457-4 · Cited by 42
“Firstly, CRISPR/Cas9 knock-in technology permits engineered T cells to express bi-allelic or sequential genes in a site-specific manner [109, 110].”
BackgroundInfluentialCappabianca, D., Pham, D. L., Forsberg, M. H., Bugel, M., Tommasi, A. M., Lauer, A. C., Vidugirienė, J., Hrdlicka, B., McHale, A., Sodji, Q. H., et al. (2024) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2024.101249 · Cited by 37
“39,41 This transitional phenotype is present in both MP and Control TRAC-CAR T cells (Figure S5).”
Zhang, D. K. Y., Brockman, J. M., Adu‐Berchie, K., Liu, Y., Binenbaum, Y., de Lázaro, I., Sobral, M. C., Tresa, R., & Mooney, D. J. (2024) · Nature Biomedical Engineering · doi:10.1038/s41551-024-01216-4 · Cited by 41
“‘Cell-extrinsic’ approaches offer key advantages over cell-intrinsic approaches, including fewer manufacturing limitations, and reduce the need for potentially detrimental modification of homoeostatic T-cell signalling networks 11 .”
Han, X., Zhang, J., Li, W., Huang, X., Wang, X., Wang, B., Gao, L., & Chen, H. (2025) · Frontiers in Immunology · doi:10.3389/fimmu.2025.1512509 · Cited by 44
Integrating T cell metabolism in cancer immunotherapy
Dugnani, E., Pasquale, V., Bordignon, C., Canu, A., Piemonti, L., & Monti, P. (2017) · Cancer Letters · doi:10.1016/j.canlet.2017.09.039 · Cited by 38
TCR repertoire evolution during maintenance of CMV ‐specific T‐cell populations
Schober, K., Buchholz, V. R., & Busch, D. H. (2018) · Immunological Reviews · doi:10.1111/imr.12654 · Cited by 43
Therapeutic gene editing in haematological disorders with CRISPR /Cas9
Jensen, T. I., Axelgaard, E., & Bak, R. O. (2019) · British Journal of Haematology · doi:10.1111/bjh.15851 · Cited by 40
“(Holkers et al, 2014; Dever et al, 2016; De Ravin et al, 2017; Eyquem et al, 2017).”
BackgroundMethodNair, R. K., & Westin, J. R. (2020) · Advances in experimental medicine and biology · doi:10.1007/978-3-030-41008-7_10 · Cited by 43
Metadata
- Authors
- Eyquem, J., Mansilla-Soto, J., Giavridis, T., van der Stegen, S. J. C., Hamieh, M., Cunanan, K. M., Odak, A., Gönen, M., & Sadelain, M.
- Affiliations
- Memorial Sloan Kettering Cancer Center (US)
- Corresponding author
- Sadelain, Michel
- ORCID
- Venue
- Nature
- Published
- 2017-02-22
- Volume
- 543
- Issue
- 7643
- Pages
- 113-117
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC5558614
- OpenAlex
- W2589417329
- Semantic Scholar
- 47af46911569825ab86ac946a5c9cd502504aa3f
- Cited by
- 1,831
- References
- 33
- Citations by year
- 2026: 191 · 2025: 175 · 2024: 206 · 2023: 201 · 2022: 224 · 2021: 238 · 2020: 190 · 2019: 186 · 2018: 151 · 2017: 68
- FWCI
- 71.74
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- Funders
- National Cancer Institute (P30CA008748)
- Topics
- Health Sciences › Medicine › Oncology › CAR-T cell therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Keywords
- chimeric antigen receptor, CRISPR/Cas9 genome editing, CD19, chimeric antigen receptor T cells, T cell exhaustion, T cell differentiation, acute lymphoblastic leukemia, tumor rejection, targeted gene integration
- MeSH terms
- Gene Editing, Animals, Cell Differentiation, Disease Models, Animal, Gene Expression Regulation, Humans, Immunotherapy, Lymphocyte Activation, Male, Promoter Regions, Genetic, Receptors, Antigen, T-Cell, T-Lymphocytes, Receptors, Antigen, T-Cell, alpha-beta, Antigens, CD19, Mice, Precursor Cell Lymphoblastic Leukemia-Lymphoma, Genetic Loci, Translational Research, Biomedical, CRISPR-Cas Systems
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