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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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“Endogenous TCR signaling also deleteriously effects CD8 CAR-T function, however, introduction of CAR sequences directly into the endogenous TCR locus may allow for appropriate tonic T cell signaling whilst minimizing the chance of random mutagenesis caused by lentiviral integration [65].”
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“A universal T cell therapy from healthy human donors must overcome multiple hurdles: The T cells must (1) be effectively reprogrammed towards the cancer via a CAR (or transgenic TCR), (2) avoid Graft-versus-Host disease (GvHD) caused by allo-reactive endogenous donor TCRs [14], and (3) be protected from immediate allo…”
Patient selection for CAR T or BiTE therapy in multiple myeloma: Which treatment for each patient?
Kegyes, D., Constantinescu, C. S., Vrancken, L., Rasche, L., Grégoire, C., Țigu, A. B., Gulei, D., Dima, D. M., Tănase, A., Einsele, H., et al. (2022) · Journal of Hematology & Oncology · doi:10.1186/s13045-022-01296-2 · Cited by 65
“Other ways of generating CAR T cells include the use of non-viral methods such as CRISPR/Cas9 [12] or transposon systems [13, 14], or more recently, in vitro-transcribed mRNA was used to reprogram T cells in vivo [15].”
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Nanomaterials for Protein Delivery in Anticancer Applications
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Self-citationProspects and challenges for use of CAR T cell therapies in solid tumors
Ramakrishna, S., Barsan, V. V., & Mackall, C. L. (2020) · Expert Opinion on Biological Therapy · doi:10.1080/14712598.2020.1738378 · Cited by 62
Advances in Developing CAR T-Cell Therapy for HIV Cure
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“Their results also showed a delay in effector T-cell differentiation and exhaustion (116).”
ResultHo, J., Wang, L., Liu, Y., Ba, M., Yang, J., Zhang, X., Chen, D., Lu, P., & Li, J. (2021) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2021.03.007 · Cited by 61
“Eyquem et al.(25) revealed that higher CAR expression by TRAC-EF1a-1928z than TRAC1928z did not produce better antitumor activity.”
BackgroundThe antigen‐binding moiety in the driver's seat of CARs
Hanssens, H., Meeus, F., Veirman, K. D., Breckpot, K., & Devoogdt, N. (2021) · Medicinal Research Reviews · doi:10.1002/med.21818 · Cited by 63
“Targeted gene delivery of a CD19‐specific CAR to the TCRα constant locus using CRISPR/Cas9 technology resulted in constant CAR expression and more potent CAR‐T cells in which tonic CAR signaling was prevented, and delayed CAR‐T cell differentiation and exhaustion was observed.(212) Altogether, fine‐tuning CAR expressi…”
Comparing CAR and TCR engineered T cell performance as a function of tumor cell exposure
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Novel strategies to manage CAR-T cell toxicity
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Engineering Induced Pluripotent Stem Cells for Cancer Immunotherapy
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“To avoid immunological rejection and minimize GvHD in patients, allogeneic T cells have to be engineered to remove their human leukocyte antigen (HLA) class I and II molecules as well as disrupt TCR expression [32,33].”
BackgroundWilson, T. L., Kim, H., Chou, C., Langfitt, D. M., Mettelman, R. C., Minervina, A. A., Allen, E. K., Métais, J., Pogorelyy, M. V., Riberdy, J. M., et al. (2022) · Cancer Discovery · doi:10.1158/2159-8290.cd-21-1508 · Cited by 58
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“, retrovirus, lentivirus): it ensures most T cells are monoclonal (only a single and unique CAR); it leads to transgene expression that is more consistent across cells and physiologically regulated; and it knocks out the endogenous TCR to avoid confounding effects [20].”
BackgroundTuned activation of MSLN-CAR T cells induces superior antitumor responses in ovarian cancer models
Schoutrop, E., Poiret, T., Serafi, I. E., Zhao, Y. H., He, R., Moter, A., Henriksson, J., Hassan, M., Magalhaes, I., & Mattsson, J. (2023) · Journal for ImmunoTherapy of Cancer · doi:10.1136/jitc-2022-005691 · Cited by 60
Managing allorejection in off-the-shelf CAR-engineered cell therapies
Li, Y.-R., Fang, Y., Niu, S., Chen, Y., Lyu, Z., & Yang, L. (2024) · Molecular Therapy · doi:10.1016/j.ymthe.2024.11.035 · Cited by 65
“21,23,84,121 However, endogenous TCRs provide certain bene fi ts for CAR-T cell function and persistence in vivo , suggesting that preserving intact TCRs may be advantageous.”
Building a Safer and Faster CAR: Seatbelts, Airbags, and CRISPR
Perales, M.-A., Kebriaei, P., Kean, L. S., & Sadelain, M. (2018) · Biology of Blood and Marrow Transplantation · doi:10.1016/j.bbmt.2017.10.017 · Cited by 59
Self-citationNext-generation cell therapies: the emerging role of CAR-NK cells
Başar, R., Daher, M., & Rezvani, K. (2020) · Hematology · doi:10.1182/hematology.2020002547 · Cited by 57
“Finally, allogeneic CAR-T cells are being explored for off-the-shelf therapy,(17,18) but given their associated risk of graft-versus-host disease (GVHD), allogeneic CAR-T cells would need further genetic modification, such as deletion of the TCR, to mitigate this risk.(19) Back to our patient Given the advanced stage…”
BackgroundOptimization and validation of CAR transduction into human primary NK cells using CRISPR and AAV
Kararoudi, M. N., Likhite, S. B., Elmas, E., Yamamoto, K., Schwartz, M., Sorathia, K., Pereira, M. D. S. F., Sezgin, Y., Devine, R. D., Lyberger, J. M., et al. (2022) · Cell Reports Methods · doi:10.1016/j.crmeth.2022.100236 · Cited by 59
“…of the flanking regions of the Cas9 targeting site in the AAVS1 locus, cloned these together with the mCherry gene into the backbone of a single-stranded AAV plasmid, and packaged this construct into the AAV6 viral capsid (Figure 2A) (Foust et al., 2013; Wang et al., 2016; Eyquem et al., 2017).”
BackgroundMethodRethinking cancer targeting strategies in the era of smart cell therapeutics
Allen, G. M., & Lim, W. A. (2022) · Nature reviews. Cancer · doi:10.1038/s41568-022-00505-x · Cited by 61
Programmable synthetic receptors: the next-generation of cell and gene therapies
Teng, F., Cui, T., Zhou, L., Gao, Q., Zhou, Q., & Li, W. (2024) · Signal Transduction and Targeted Therapy · doi:10.1038/s41392-023-01680-5 · Cited by 70
CRISPR–Cas9 delivery strategies for the modulation of immune and non-immune cells
Alsaiari, S. K., Eshaghi, B., Du, B., Kanelli, M., Li, G., Wu, X., Zhang, L., Chaddah, M., Lau, A., Yang, X., et al. (2024) · Nature Reviews Materials · doi:10.1038/s41578-024-00725-7 · Cited by 63
Esmaeilzadeh, A., Tahmasebi, S., & Athari, S. S. (2019) · Biomedicine & Pharmacotherapy · doi:10.1016/j.biopha.2019.109685 · Cited by 70
Regulatory T cell therapy: Current and future design perspectives
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T Cell-Redirecting Strategies to ‘STAb’ Tumors: Beyond CARs and Bispecific Antibodies
Blanco, B., Compte, M., Lykkemark, S., Sanz, L., & Álvarez‐Vallina, L. (2019) · Trends in Immunology · doi:10.1016/j.it.2019.01.008 · Cited by 59
Xu, X., Huang, S., Xiao, X., Sun, Q., Liang, X., Chen, S., Zhao, Z., Huo, Z., Tu, S., & Li, Y. (2021) · Frontiers in Immunology · doi:10.3389/fimmu.2020.569117 · Cited by 62
“Using an automation system (CliniMACS Prodigy System) (132), nonviral transposon-based systems (133–135), a type II CRISPR/Cas9 system (136) and transferring CAR coding sequences in vivo (137, 138) could reduce the manufacturing time and cost as well as manifest the high potency and proliferation capacity of CAR T-cel…”
MethodEmerging CAR T Cell Strategies for the Treatment of AML
Vishwasrao, P., Li, G., Boucher, J. C., Smith, D. L., & Hui, S. K. (2022) · Cancers · doi:10.3390/cancers14051241 · Cited by 61
A review on CRISPR/Cas: a versatile tool for cancer screening, diagnosis, and clinic treatment
Yang, X., & Zhang, B. (2023) · Functional & Integrative Genomics · doi:10.1007/s10142-023-01117-w · Cited by 58
“Eyquem and colleagues employed CRISPR/Cas9 genome editing to successfully knock in CAR DNA cassette into the genome site of endogenous T-cell receptor α constant (TRAC) locus; the CRISPR-generated CAR T-cell not only knock in CAR DNA sequences but also knock out TRAC gene (Eyquem et al. 2017; Kath et al. 2022).”
BackgroundInfluentialChimeric antigen receptor engineered natural killer cells for cancer therapy
Zhang, Y., Zhou, W., Yang, J., Yang, J., & Wang, W. (2023) · Experimental Hematology and Oncology · doi:10.1186/s40164-023-00431-0 · Cited by 58
“Directing CD19 CAR to the T-cell receptor α constant (TRAC) locus by CRISPR/ Cas9 resulted in consistent CAR expression in human peripheral blood T cells as well as improved effector responses [293].”
BackgroundCRISPR and personalized Treg therapy: new insights into the treatment of rheumatoid arthritis
Safari, F., Farajnia, S., Aria, M., Zarredar, H., & Nasrolahi, A. (2018) · Immunopharmacology and Immunotoxicology · doi:10.1080/08923973.2018.1437625 · Cited by 58
Genetic Modification Strategies to Enhance CAR T Cell Persistence for Patients With Solid Tumors
DeRenzo, C. C., & Gottschalk, S. M. (2019) · Frontiers in Immunology · doi:10.3389/fimmu.2019.00218 · Cited by 57
“Another promising strategy is to utilize targeted gene editing and insert CAR constructs into T cell inhibitory loci (66, 67).”
BackgroundExpanding the CRISPR Toolbox with ErCas12a in Zebrafish and Human Cells
Wierson, W. A., Simone, B. W., WareJoncas, Z., Mann, C. M., Welker, J. M., Kar, B., Emch, M. J., Friedberg, I., Gendron, W. A. C., Barry, M. A., et al. (2019) · The CRISPR Journal · doi:10.1089/crispr.2019.0026 · Cited by 57
“4E).(29) In vitro cleavage activity of ErCas12a with our expression system was assessed using a T7 endonuclease (T7EI) assay to determine whether a DSB and subsequent indels were induced at the AAVS1 locus after transfection into HEK293T cells.”
MethodMolecular Imaging with Reporter Genes: Has Its Promise Been Delivered?
Serganova, I. S., & Blasberg, R. G. (2019) · Journal of Nuclear Medicine · doi:10.2967/jnumed.118.220004 · Cited by 60
“The first is the need to transduce cells and target tissues with specific reporter constructs, and the second is the absence of optimal vectors for targeting specific organs or tissues, although considerable progress has been made in the ex vivo transduction of human-derived cells (227,231,232) and this continues to b…”
BackgroundProgress and challenges towards CRISPR/Cas clinical translation
Rosenblum, D., Gutkin, A., Dammes, N., & Peer, D. (2020) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2020.07.004 · Cited by 61
Targeted T cell receptor gene editing provides predictable T cell product function for immunotherapy
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“In the future, more long-term in vivo protection experiments are needed to test whether OTR T cells show prolonged maintenance and enhanced effector function, as proposed by more physiological TCR regulation(32) and OTR CAR-T cell data.(44) We observed by scRNAseq aswell as flow cytometry that OTR decreases cell-to-ce…”
Innate and Innate-Like Cells: The Future of Chimeric Antigen Receptor (CAR) Cell Therapy
Cortés‐Selva, D., Dasgupta, B., Singh, S., & Grewal, I. S. (2020) · Trends in Pharmacological Sciences · doi:10.1016/j.tips.2020.11.004 · Cited by 61
From TCR fundamental research to innovative chimeric antigen receptor design
Minguet, S., Maus, M. V., & Schamel, W. W. A. (2024) · Nature reviews. Immunology · doi:10.1038/s41577-024-01093-7 · Cited by 58
“In addition to serial triggering, the high affinity of CARs for their ligands may alter receptor recycling, signalling and mechanoreceptor function, properties that are thought to contribute to the high sensitivity of TCRs 87,160 .”
Addressing graft-versus-host disease in allogeneic cell-based immunotherapy for cancer
Lyu, Z., Niu, S., Fang, Y., Chen, Y., Li, Y.-R., & Yang, L. (2025) · Experimental Hematology and Oncology · doi:10.1186/s40164-025-00654-3 · Cited by 61
“This approach enables the targeted recognition of specific antigens, enhancing the safety and effectiveness of these cells for therapeutic applications, as demonstrated in several preclinical studies [17, 78, 113, 114].”
InfluentialHierarchical Micro‐/Nanostructures from Human Hair for Biomedical Applications
Zheng, D., Hong, S., Xu, L., Li, C., Li, K., Cheng, S., & Zhang, X. (2018) · Advanced Materials · doi:10.1002/adma.201800836 · Cited by 57
CRISPR-Based Tools in Immunity
Simeonov, D. R., & Marson, A. (2019) · Annual Review of Immunology · doi:10.1146/annurev-immunol-042718-041522 · Cited by 57
The potential of CAR T cell therapy for prostate cancer
Wolf, P., Alzubi, J., Gratzke, C., & Cathomen, T. (2021) · Nature Reviews Urology · doi:10.1038/s41585-021-00488-8 · Cited by 58
Drug-regulated CD33-targeted CAR T cells control AML using clinically optimized rapamycin dosing
Appelbaum, J., Price, A. E., Oda, K., Zhang, J., Leung, W., Tampella, G., Xia, D., So, P. P., Hilton, S. K., Evandy, C., et al. (2024) · Journal of Clinical Investigation · doi:10.1172/jci162593 · Cited by 56
“In addition, periods of alternating signaling and quiescence may enhance the efficacy of antitumor T cells by preventing T cell exhaustion (48–50) and allowing effector T cells to transition to memory states (29, 30, 51–54) after periods of prolonged antigen exposure.”
BackgroundCRISPR/Cas9 genome editing: Fueling the revolution in cancer immunotherapy
Liu, X., & Zhao, Y. (2018) · Current Research in Translational Medicine · doi:10.1016/j.retram.2018.04.003 · Cited by 57
Chimeric antigen receptor T cells in solid tumors: a war against the tumor microenvironment
Zhao, Z., Xiao, X., Saw, P. E., Wu, W., Huang, H., Chen, J., & Nie, Y. (2019) · Science China Life Sciences · doi:10.1007/s11427-019-9665-8 · Cited by 56
Strategies for Dodging the Obstacles in CAR T Cell Therapy
Kozani, P. S., Kozani, P. S., Rahbarizadeh, F., & Nikkhoi, S. K. (2021) · Frontiers in Oncology · doi:10.3389/fonc.2021.627549 · Cited by 57
Hoerster, K., Uhrberg, M., Wiek, C., Horn, P. A., Hanenberg, H., & Heinrichs, S. (2021) · Frontiers in Immunology · doi:10.3389/fimmu.2020.586168 · Cited by 59
“An interesting approach to abrogate unwanted or alloreactive signaling from the endogenous T cell receptors (TCRs) in chimeric antigen receptor (CAR) T cells uses genome editing on common TCR domains (12, 13), however this genomic editing will require additional gene transfer systems and therefore will add several lay…”
BackgroundOu, X., Ma, Q., Yin, W., Ma, X., & He, Z. (2021) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2021.674467 · Cited by 58
Engineering cells for therapy and diagnosis
Li, Z., Wang, Y., Gu, Z., & Hu, Q. (2024) · Nature Reviews Bioengineering · doi:10.1038/s44222-024-00198-x · Cited by 60
“from healthy donors can be used to prepare allogeneic CAR T cells, thereby extending the application scope of CAR T cell therapy 29 .”
Acute lymphoblastic leukemia relapse after CD19-targeted chimeric antigen receptor T cell therapy
Wang, J., Hu, Y., & Huang, H. (2017) · Journal of Leukocyte Biology · doi:10.1189/jlb.5ru0817-315r · Cited by 57
“[40] found that the surface CAR level was negatively associated with tumor burden as a result of the internalization of the receptors.”
BackgroundLee, J., Sheen, J. H., Lim, O., Lee, Y. J., Ryu, J., Shin, D., Kim, Y. Y., & Kim, M. (2020) · Scientific Reports · doi:10.1038/s41598-020-74772-9 · Cited by 54
Ruggiero, E., Carnevale, E., Prodeus, A., Magnani, Z. I., Camisa, B., Merelli, I., Politano, C., Stasi, L., Potenza, A., Cianciotti, B. C., et al. (2022) · Science Translational Medicine · doi:10.1126/scitranslmed.abg8027 · Cited by 58
CRISPR–Cas9 applications in T cells and adoptive T cell therapies
Chen, X., Zhong, S., Zhan, Y., & Zhang, X. (2024) · Cellular & Molecular Biology Letters · doi:10.1186/s11658-024-00561-1 · Cited by 58
“CRISPR–Cas9 makes it accurately insert the CAR into the TRAC locus to enhance its anticancer activity and overcome the random vector integration challenges [120, 123].”
BackgroundRetracted: Applications and advances of CRISPR-Cas9 in cancer immunotherapy
Xia, A., He, Q., Wang, J., Zhu, J., Sha, Y., Sun, B., & Lu, X. (2018) · Journal of Medical Genetics · doi:10.1136/jmedgenet-2018-105422 · Cited by 57
RetractedChimeric Antigen Receptor Library Screening Using a Novel NF-κB/NFAT Reporter Cell Platform
Rydzek, J., Nerreter, T., Peng, H., Jutz, S., Leitner, J. M., Steinberger, P., Einsele, H., Rader, C. M., & Hudecek, M. (2018) · Molecular Therapy · doi:10.1016/j.ymthe.2018.11.015 · Cited by 56
Promoter choice: Who should drive the CAR in T cells?
Rad, S. M. A. H., Poudel, A., Tan, G. M. Y., & McLellan, A. D. (2020) · PLoS ONE · doi:10.1371/journal.pone.0232915 · Cited by 62
“However, the effects of promoter silencing might be overshadowed by the long term CAR T cell downregulation that occurs in a methylation-independent fashion following CAR triggering both in vitro and in vivo [14, 41, 42].”
BackgroundEmerging Strategies in TCR-Engineered T Cells
Fang, W., Cheng, X., Xue, J. Z., & Xue, S. (2022) · Frontiers in Immunology · doi:10.3389/fimmu.2022.850358 · Cited by 58
“Several strategies have been explored to achieve this goal, including the use of siRNAs (66, 67), zinc-fi nger nucleases (68), transcription activator-like effector nucleases (TALENs) (69, 70), and CRISPR/Cas9 technologies (71, 72).”
BackgroundMethodModified Hypoxia-Inducible Factor Expression in CD8+ T Cells Increases Antitumor Efficacy
Veliça, P., Cunha, P. P., Vojnovic, N., Foskolou, I. P., Bargiela, D., Gojkovic, M., Rundqvist, H., & Johnson, R. S. (2021) · Cancer Immunology Research · doi:10.1158/2326-6066.cir-20-0561 · Cited by 52
Evolution of CD8+ T Cell Receptor (TCR) Engineered Therapies for the Treatment of Cancer
Sun, Y., Li, F., Sonnemann, H. M., Jackson, K. R., Talukder, A. H., Katailiha, A. S., & Lizee, G. A. (2021) · Cells · doi:10.3390/cells10092379 · Cited by 56
“Genome editing has brought the promise of specific and efficient insertions of large transgenes into target cells through homology-directed repair (HDR) [65,66].”
BackgroundMassively parallel knock-in engineering of human T cells
Dai, X., Park, J. J.-Y., Du, Y., Na, Z., Lam, S. Z., Chow, R. D.-W., Renauer, P. A., Gu, J., Shan, X., Chu, Z., et al. (2023) · Nature Biotechnology · doi:10.1038/s41587-022-01639-x · Cited by 56
CRISPR/Cas-based CAR-T cells: production and application
Song, P., Zhang, Q., Xu, Z., Shi, Y., Jing, R., & Luo, D. (2024) · Biomarker Research · doi:10.1186/s40364-024-00602-z · Cited by 55
“In comparison to CAR-T cells infected with retroviral vectors, the CD19 CAR knockin CAR-T cells exhibited diminished differentiation and depletion, while demonstrating significantly improved anti-tumor effects in mouse models [16].”
BackgroundMethodResultInfluentialImmunoengineering with biomaterials for enhanced cancer immunotherapy
Xie, Y., Wei, L., & Tang, L. (2018) · Wiley Interdisciplinary Reviews Nanomedicine and Nanobiotechnology · doi:10.1002/wnan.1506 · Cited by 53
Gut microbiome and CAR-T therapy
Abid, M. B., Shah, N. N., Maatman, T. C., & Hari, P. (2019) · Experimental Hematology and Oncology · doi:10.1186/s40164-019-0155-8 · Cited by 57
CRISPR technology: The engine that drives cancer therapy
Zhang, M., Eshraghian, E. A., Jammal, O. A., Zhang, Z.-B., & Zhu, X. M. (2020) · Biomedicine & Pharmacotherapy · doi:10.1016/j.biopha.2020.111007 · Cited by 54
Automated generation of gene-edited CAR T cells at clinical scale
Alzubi, J., Lock, D., Rhiel, M., Schmitz, S., Wild, S., Mussolino, C., Hildenbeutel, M., Brandes, C., Rositzka, J., Lennartz, S., et al. (2021) · Molecular Therapy - Methods & Clinical Development · doi:10.1016/j.omtm.2020.12.008 · Cited by 53
Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins
Kararoudi, M. N., Dolatshad, H., Trikha, P., Hussain, S.-R. A., Elmas, E., Foltz, J. A., Moseman, J. E., Thakkar, A., Nakkula, R. J., Lamb, M., et al. (2018) · Journal of Visualized Experiments · doi:10.3791/58237 · Cited by 64
Optimized CRISPR-mediated gene knockin reveals FOXP3-independent maintenance of human Treg identity
Lam, A. J., Lin, D. T. S., Gillies, J. K., Uday, P., Pesenacker, A. M., Kobor, M. S., & Levings, M. K. (2021) · Cell Reports · doi:10.1016/j.celrep.2021.109494 · Cited by 52
“CRISPR-mediated HDR has been used in human T cells to correct point mutations or replace the endogenous TCR with an engineered TCR or chimeric antigen receptor (Eyquem et al., 2017; MacLeod et al., 2017; Roth et al., 2018).”
InfluentialThe Progress and Prospects of Immune Cell Therapy for the Treatment of Cancer
Han, J., Zhang, B., Zheng, S., Jiang, Y. H., Zhang, X., & Mao, K. (2024) · Cell Transplantation · doi:10.1177/09636897241231892 · Cited by 57
“Compared with random integration, CAR expression can be regulated by the endogenous TCR promoter and mimics TCR transcription when exposed to antigen, preventing constant, and excessive T cell activation, which could result in T cell differentiation and exhaustion 97 .”
BackgroundMethodResultInfluentialCRISPR-Mediated Editing of the B Cell Receptor in Primary Human B Cells
Greiner, V., Bou-Puerto, R., Liu, S., Herbel, C., Carmona, E. M., & Goldberg, M. S. (2019) · iScience · doi:10.1016/j.isci.2019.01.032 · Cited by 54
“Clinical translation of genome-edited cellular therapies, whose function and safety can be controlled through synthetic biology (Fischbach et al., 2013), heralds an emerging era in the treatment of genetic, infectious, and acquired diseases (Cornu et al., 2017).”
BackgroundEffects of pericytes and colon cancer stem cells in the tumor microenvironment
Garza-Treviño, E. N., González, P. D., Salgado, C. I. V., & Garza, A. M. (2019) · Cancer Cell International · doi:10.1186/s12935-019-0888-9 · Cited by 53
All systems go: converging synthetic biology and combinatorial treatment for CAR-T cell therapy
Lanitis, E., Coukos, G., & Irving, M. B. (2020) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2020.01.009 · Cited by 52
Breaking Bottlenecks for the TCR Therapy of Cancer
Gaissmaier, L., Elshiaty, M., & Christopoulos, P. (2020) · Cells · doi:10.3390/cells9092095 · Cited by 52
Yi, Y., Chai, X., Zheng, L., Zhang, Y., Shen, J.-K., Hu, B., & Tao, G. (2021) · Cell Discovery · doi:10.1038/s41421-021-00255-4 · Cited by 54
Sudarsanam, H., Buhmann, R., & Henschler, R. (2022) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2022.886637 · Cited by 54
Integration of ζ-deficient CARs into the CD3 ζ gene conveys potent cytotoxicity in T and NK cells
Kath, J., Franke, C., Drosdek, V., Du, W., Glaser, V., Fuster‐García, C., Stein, M., Zittel, T., Schulenberg, S., Porter, C. E., et al. (2024) · Blood · doi:10.1182/blood.2023020973 · Cited by 52
CD5 deletion enhances the antitumor activity of adoptive T cell therapies
Patel, R. P., Ghilardi, G., Zhang, Y., Chiang, Y., Xie, W., Guruprasad, P., Kim, K. H., Chun, I., Angelos, M. G., Pajarillo, R., et al. (2024) · Science Immunology · doi:10.1126/sciimmunol.adn6509 · Cited by 55
“To this goal, we generated TCR α constant (TRAC) KO T cells (29) further transduced with TCR-GP100 lentivirus (30) and compared these to cells with a double KO (DKO) of both TRAC and CD5 using the GP100 + melanoma cell line DM6.”
CRISPR-Cas9: a promising genetic engineering approach in cancer research
Ratan, Z. A., Son, Y., Haidere, M. F., Uddin, B. M. M., Yusuf, M. A., Zaman, S. B., Kim, J., Banu, L. A., & Cho, J. Y. (2018) · Therapeutic Advances in Medical Oncology · doi:10.1177/1758834018755089 · Cited by 52
Chimeric antigen receptor (CAR)-modified NK cells against cancer: Opportunities and challenges
Wang, L., Dou, M., Ma, Q., Yao, R., & Liu, J. (2019) · International Immunopharmacology · doi:10.1016/j.intimp.2019.105695 · Cited by 51
CAR- and TRuC-redirected regulatory T cells differ in capacity to control adaptive immunity to FVIII
Rana, J., Perry, D. J., Kumar, S. R. P., Muñoz-Melero, M., Saboungi, R., Brusko, T. M., & Biswas, M. (2021) · Molecular Therapy · doi:10.1016/j.ymthe.2021.04.034 · Cited by 51
“A related study targeting a CAR to the TRAC locus was shown to avert tonic signaling in a mouse model of acute lymphoblastic leukemia by a mechanism of CAR internalization and post-stimulation replenishment of cell-surface CAR expression.(66) TRuCs employ the entire TCR complex to signal, whereas CARs utilize only the…”
BackgroundNK Cells Armed with Chimeric Antigen Receptors (CAR): Roadblocks to Successful Development
Dezfouli, A. B., Yazdi, M., Pockley, A. G., Khosravi, M., Kobold, S., Wagner, E., & Multhoff, G. (2021) · Cells · doi:10.3390/cells10123390 · Cited by 53
Cook, M. S., King, E., Flaherty, K. R., Siddika, K., Papa, S., Benjamin, R., & Schurich, A. (2025) · Cell Reports · doi:10.1016/j.celrep.2025.115973 · Cited by 50
Clinical applications of CRISPR‐based genome editing and diagnostics
Foss, D. V., Hochstrasser, M. L., & Wilson, R. C. (2019) · Transfusion · doi:10.1111/trf.15126 · Cited by 51
Engineering γδT cells limits tonic signaling associated with chimeric antigen receptors
Fisher, J. P. H., Sharma, R., Don, D. W., Barisa, M., Hurtado, M. O., Abramowski, P., Porter, L., Day, W. P., Borea, R., Inglott, S., et al. (2019) · Science Signaling · doi:10.1126/scisignal.aax1872 · Cited by 51
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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