The DOI matches its Crossref record; 33 of the 33 references it cites were found in a registry or an index.
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.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“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
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
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
Cited by
Sorted by citations
Items 601–700 of 1,002
Huang, D., Miller, M., Ashok, B., Jain, S., & Peppas, N. A. (2020) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2020.07.015 · Cited by 38
“This strategy also further minimizes risks of insertional oncogenesis and TCR-induced autoimmunity for universal allogeneic T cells [58,59].”
BackgroundMethodTargeted Gene Delivery: Where to Land
Pavani, G., & Amendola, M. (2021) · Frontiers in Genome Editing · doi:10.3389/fgeed.2020.609650 · Cited by 41
“New CAR-T cells are generated by targeting the integration of the CAR transgene under the transcriptional control of TCR α-chain gene promoter to simultaneously achieve physiological expression of CAR and disruption of the endogenous TCR, thus maintaining only CAR antigen specificity (specificity exchange) (Eyquem et…”
BackgroundSynthetic TILs: Engineered Tumor-Infiltrating Lymphocytes With Improved Therapeutic Potential
Jiménez-Reinoso, A., Nehme-Álvarez, D., Domínguez-Alonso, C., & Álvarez‐Vallina, L. (2021) · Frontiers in Oncology · doi:10.3389/fonc.2020.593848 · Cited by 38
“Engineered TCR- and CAR-T cells are leukapheresis blood-derived T cells genetically modified ex vivo in order to specifically recognize a tumor-associated antigen (TAA) viamRNA electroporation (16), lentiviral (17) or retroviral (18) transduction, transposon mediated modification (19) or via CRISPR/Cas9 gene editing (…”
BackgroundGlycoprotein Targeted CAR-NK Cells for the Treatment of SARS-CoV-2 Infection
Christodoulou, I., Rahnama, R., Ravich, J. W., Seo, J., Zolov, S. N., Marple, A., Markovitz, D. M., & Bonifant, C. L. (2021) · Frontiers in Immunology · doi:10.3389/fimmu.2021.763460 · Cited by 38
“We observed constitutive CD3z phosphorylation of the CAR in our transduced cells, a finding that has previously been associated with CAR tonic signaling (Figure 1C) (37, 38).”
BackgroundEngineered T Cells: CAR T Cell Therapy and Beyond
Johnson, P. C., & Abramson, J. S. (2022) · Current Oncology Reports · doi:10.1007/s11912-021-01161-4 · Cited by 39
“Interestingly, targeting the CAR to the TRAC locus also averted tonic signaling and allowed for internalization and re-expression of the CAR with repeated antigen exposure, delaying effector T cell differentiation and ultimately preventing T cell exhaustion [34].”
BackgroundChimeric Antigen Receptor (CAR) Regulatory T-Cells in Solid Organ Transplantation
Gille, I., Claas, F. H. J., Haasnoot, G. W., Heemskerk, M. H. M., & Heidt, S. (2022) · Frontiers in Immunology · doi:10.3389/fimmu.2022.874157 · Cited by 40
“By disrupting the TRAC locus, endogenous TCRs are not expressed anymore and in addition, this will lead to uniform CAR expression and enhanced potency in T-cells (93).”
BackgroundMakita, S., Imaizumi, K., Kurosawa, S., & Tobinai, K. (2019) · Drugs in Context · doi:10.7573/dic.212567 · Cited by 40
Ding, J. P., Guyette, S., Schrand, B., Geirut, J., Horton, H. M., Guo, G., Delgoffe, G. M., Menk, A. V., Baeuerle, P. A., Hofmeister, R., et al. (2023) · OncoImmunology · doi:10.1080/2162402x.2023.2182058 · Cited by 35
“Such differential transgene expression may be because TRuCs are less prone to downregulation than CARs upon antigen encounter.(20,21) Interestingly, TC-210 T cells were more efficient in controlling early tumor growth than CAR-T cells despite releasing a”
BackgroundNg, B. D., Rajagopalan, A., Kousa, A. I., Fischman, J. S., Chen, S., Massa, A., Elias, H. K., Manuele, D., Galiano, M., Lemarquis, A. L., et al. (2024) · Blood · doi:10.1182/blood.2023022293 · Cited by 40
Dimitri, A. J., Baxter, A. E., Chen, G. M., Hopkins, C. R., Rouin, G. T., Huang, H., Kong, W., Holliday, C., Wiebking, V., Bartoszek, R. L., et al. (2024) · Science Advances · doi:10.1126/sciadv.adp9371 · Cited by 42
“This approach was designed to delay effector T cell differentiation and exhaustion through CAR insertion at the TRAC locus as previously described ( 67 ), while leveraging the potential benefits of 41BB costimulation ( 4 ).”
Tracing the development of CAR-T cell design: from concept to next-generation platforms
Alsaieedi, A., & Zaher, K. A. (2025) · Frontiers in Immunology · doi:10.3389/fimmu.2025.1615212 · Cited by 49
Toward “off‐the‐shelf” allogeneic CAR T cells
Aftab, B. T., Sasu, B. J., Krishnamurthy, J., Gschweng, E. H., Alcazer, V., & Depil, S. (2020) · Advances in Cell and Gene Therapy · doi:10.1002/acg2.86 · Cited by 40
Odé, Z., Condori, J., Peterson, N. B., Zhou, S., & Krenciute, G. (2020) · Cancers · doi:10.3390/cancers12061704 · Cited by 37
“Based on these rules, our final donor DNA for the TRAC locus contained the following parts: 400 bp LHA, spliced acceptor (SA) [18], P2A (porcine teschovirus-1 2A peptide), IL-15, E2A (equine rhinitis A virus 2A peptide), mClover3, poly(A) (bovine growth hormone polyadenylation signal), and 400 bp RHA (Figure 2c).”
BackgroundMethodInfluentialEnhancing site-specific DNA integration by a Cas9 nuclease fused with a DNA donor-binding domain
Ma, S., Wang, X., Hu, Y., Lv, J., Liu, C., Liao, K., Guo, X., Wang, D., Lin, Y., & Rong, Z. (2020) · Nucleic Acids Research · doi:10.1093/nar/gkaa779 · Cited by 36
Ponterio, E., Maria, R. D., & Haas, T. L. (2020) · Frontiers in Immunology · doi:10.3389/fimmu.2020.565631 · Cited by 37
“Other ways to enhance the safety of CARs recognizing TAAs also expressed by healthy tissues is a stringent control of their expression levels either by transient expression methods usingmRNA transfer instead of stable transduction (81) or the integration of the transgene in a defined genetic locus enabling controlled…”
MethodImmune cell labelling and tracking: implications for adoptive cell transfer therapies
Galli, F., Varani, M., Lauri, C., Silveri, G. G., Onofrio, L., & Signore, A. (2021) · EJNMMI Radiopharmacy and Chemistry · doi:10.1186/s41181-020-00116-7 · Cited by 37
Donor T cells for CAR T cell therapy
Tang, T. C. Y., Xu, N., Nordon, R. E., Haber, M., Micklethwaite, K. P., & Dolnikov, A. (2022) · Biomarker Research · doi:10.1186/s40364-022-00359-3 · Cited by 43
Gene therapy for blood diseases
Kohn, D. B. (2019) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2018.11.016 · Cited by 39
Function and evolution of the prototypic CD28ζ and 4-1BBζ chimeric antigen receptors
Feucht, J., & Sadelain, M. (2020) · Immuno-Oncology Technology · doi:10.1016/j.iotech.2020.09.001 · Cited by 42
“TRAC-CAR T cells exhibit reduced tonic signalling, delayed T-cell exhaustion, differentiation and enhanced antitumour activity relative to conventionally engineered CAR T cells.(66) Similarly, TRAC-19BBz CAR T cells yield constant CAR expression and achieve superior tumour eradication in vivo.”
BackgroundInfluentialSelf-citationAdvances in CRISPR Delivery Methods: Perspectives and Challenges
Demirci, S., Essawi, K., Germino-Watnick, P., Liu, X., Hakami, W., & Tisdale, J. F. (2022) · The CRISPR Journal · doi:10.1089/crispr.2022.0051 · Cited by 37
“87 These findings suggest the applicability and efficiency of AAV as a vehicle to deliver genome editing tools.”
Factors associated with long-term outcomes of CD19 CAR T-cell therapy for relapsed/refractory CLL
Liang, E. C., Albittar, A. A., Huang, J. J., Hirayama, A. V. V., Kimble, E. L., Portuguese, A. J., Chapuis, A. G., Shadman, M. S., Till, B. G., Cassaday, R. D., et al. (2023) · Blood Advances · doi:10.1182/bloodadvances.2023011399 · Cited by 38
“22-24 Strategies to bolster CAR T-cell function and persistence include improving the CAR construct, for example by targeting the CD19 CAR to the T-cell receptor α constant locus 25 or using a fully human CD19 binder, 26,27 or combinatorial strategies such as concurrent PD-L1 blockade.”
BackgroundRationally designed approaches to augment CAR-T therapy for solid tumor treatment
Zhu, C., Wu, Q., Sheng, T., Shi, J., Shen, X., Yu, J., Du, Y., Sun, J., Liang, T., He, K., et al. (2023) · Bioactive Materials · doi:10.1016/j.bioactmat.2023.11.002 · Cited by 39
“The current solution is to knock down MHC-I complex and T cell receptors in allogeneic T cells via genetic engineering tools like zinc finger nucleases [135], transcription activator-like effector nuclease [136], and CRISPR/Cas9 [137].”
Chen, X., Tan, B., Xing, H., Zhao, X., Ping, Y., Zhang, Z., Huang, J., Shi, X., Zhang, N., Lin, B., et al. (2024) · Cancer Immunology Immunotherapy · doi:10.1007/s00262-023-03586-1 · Cited by 42
“Transcription activator-like effector nuclease (TALEN) or CRISPR/Cas9-edited TRAC for CAR-T19 cells exhibited antitumor immune responses in clinicals [32, 33].”
BackgroundXie, K., Starzyk, J., Majumdar, I., Wang, J., Rincones, K., Tran, T. T. D., Lee, D., Niemi, S., Famiglietti, J., Suter, B., et al. (2024) · Nature Biotechnology · doi:10.1038/s41587-024-02504-9 · Cited by 36
“Targeting a CAR to the TRAC locus greatly enhanced the potency of CAR-T cells in preclinical studies and delayed effector T cell differentiation and exhaustion 1 .”
Qin, H., Zhou, Z., Shi, R., Mai, Y., Xu, Y., Fu, P., Cheng, G., Zhang, Z., Chen, W., Chen, Y., et al. (2025) · Journal of Hematology & Oncology · doi:10.1186/s13045-025-01701-6 · Cited by 43
Highly Efficient and Versatile Plasmid-Based Gene Editing in Primary T Cells
Kornete, M., Marone, R., & Jeker, L. T. (2018) · The Journal of Immunology · doi:10.4049/jimmunol.1701121 · Cited by 33
“Targeted insertion of a CAR into the endogenous TCR locus is likely safer and results in more efficient CAR T cells compared with virally delivered randomly integrated CAR constructs (7, 34).”
BackgroundHan, C., Sim, S.-J., Kim, S.-H., Singh, R., Hwang, S., Kim, Y. I., Park, S. H., Kim, K. H., Lee, D. G., Oh, H. S., et al. (2018) · Nature Communications · doi:10.1038/s41467-018-02912-x · Cited by 37
Daniel-Moreno, A., Lamsfus‐Calle, A., Raju, J., Antony, J. S., Handgretinger, R., & Mezger, M. (2019) · Bone Marrow Transplantation · doi:10.1038/s41409-019-0510-8 · Cited by 40
A long way to the battlefront: CAR T cell therapy against solid cancers
Wang, Z., Chen, W., Zhang, X., Cai, Z., & Huang, W. (2019) · Journal of Cancer · doi:10.7150/jca.30406 · Cited by 37
“CAR T cell activation is also affected by the expression of CARs on the cell surface [35].”
BackgroundCAR-expressing NK cells for cancer therapy: a new hope
Xia, J., Minamino, S., & Kuwabara, K. (2020) · BioScience Trends · doi:10.5582/bst.2020.03308 · Cited by 36
“A few studies have suggested that editing a CAR by using the CRISPR/Cas9 technique could cause homogeneous CAR expression and improve cytotoxicity efficiency (59).”
BackgroundSingh, D. D., Han, I., Choi, E. H., & Yadav, D. K. (2021) · Computational and Structural Biotechnology Journal · doi:10.1016/j.csbj.2021.04.036 · Cited by 39
“CRISPR/Cas9 technology was employed to knock out PDCD1, TRAC, and beta-2microglobulin (b2M), which encodes the accessory chain of MHC class I in CD19 or PSCA CAR T cells [162].”
BackgroundMethodInfluentialCas9 RNP transfection by vapor nanobubble photoporation for ex vivo cell engineering
Raes, L., Pille, M., Harizaj, A., Goetgeluk, G., Hoeck, J. V., Stremersch, S., Fraire, J. C., Brans, T., de Jong, O. G., Maas-Bakker, R. F., et al. (2021) · Molecular Therapy — Nucleic Acids · doi:10.1016/j.omtn.2021.08.014 · Cited by 38
“Genome-editing strategies, such as CRISPR-Cas9, have also been extensively investigated in recent years to enhance the efficacy of next-generation CAR T cell therapeutics.(51,52) Indeed, CAR T cell activity is often insufficient to invoke robust tumor clearance as a result of tumor-induced immunosuppression.”
Emerging strategies for treating autoimmune disorders with genetically modified Treg cells
Boardman, D. A., & Levings, M. K. (2022) · Journal of Allergy and Clinical Immunology · doi:10.1016/j.jaci.2021.11.007 · Cited by 36
Engineered NK Cells Against Cancer and Their Potential Applications Beyond
Karvouni, M., Vidal-Manrique, M., Lundqvist, A., & Alici, E. (2022) · Frontiers in Immunology · doi:10.3389/fimmu.2022.825979 · Cited by 38
Current advances of CRISPR-Cas technology in cell therapy
Qiu, H.-Y., Ji, R.-J., & Zhang, Y. (2022) · Cell Insight · doi:10.1016/j.cellin.2022.100067 · Cited by 38
“Justin Eyquem succeeded in combing electroporation of Cas9 mRNA and sgRNA with rAAV6 transduction carrying CAR cDNA to precisely insert CAR into the T-cell receptor α constant (TRAC) locus (Eyquem et al., 2017).”
MethodMorton, L. T., Wachsmann, T. L. A., Meeuwsen, M. H., Wouters, A. K., Remst, D. F. G., van Loenen, M. M., Falkenburg, J. H. F., & Heemskerk, M. H. M. (2022) · Journal for ImmunoTherapy of Cancer · doi:10.1136/jitc-2021-003715 · Cited by 35
Advances in modular control of CAR-T therapy with adapter-mediated CARs
McCue, A. C., Yao, Z., & Kuhlman, B. A. (2022) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2022.114358 · Cited by 38
Zheng, H., Chen, Y., Luo, Q., Zhang, J., Huang, M., Xu, Y., Huo, D., Shan, W., Tie, R., Zhang, M., et al. (2023) · Cell Regeneration · doi:10.1186/s13619-023-00175-6 · Cited by 34
“…some approaches making CAR-T cells more persistent and enhanced anti-tumor activity in mouse tumor models, like inhibiting TCR-CD3 complex, CD155, β2-microglobulin, and EZH1, or overexpressing IL-15, IL-15R, and antigen E (Eyquem et al. 2017; Jing et al. 2022; Ueda et al. 2023; Wang et al. 2021).”
BackgroundDegagné, É., Donohoue, P. D., Roy, S., Scherer, J., Fowler, T. W., Davis, R. T., Reyes, G. A., Kwong, G., Stanaway, M., Vicena, V. L., et al. (2024) · Cancer Immunology Research · doi:10.1158/2326-6066.cir-23-0679 · Cited by 35
“Many designs had editing ef fi ciencies equivalent to an all-RNA crRNA, with chRDNA designs with DNA bases at positions 1, 11, and 20 in the spacer being among the top designs emerging from the screens.”
Directed evolution of hyperactive integrases for site specific insertion of transgenes
Hew, B. E., Gupta, S., Sato, R., Waller, D. F., Stoytchev, I., Short, J. E., Sharek, L., Tran, C. T., Badran, A. H., & Owens, J. B. (2024) · Nucleic Acids Research · doi:10.1093/nar/gkae534 · Cited by 35
Unlocking the potential of engineered immune cell therapy for solid tumors
Albarrán, V., Angelats, L., Delgado, J., Gros, A., Urbano-Ispizúa, Á., Guedan, S., & Prat, A. (2025) · Nature Communications · doi:10.1038/s41467-025-56527-0 · Cited by 39
Gene regulation technologies for gene and cell therapy
Butterfield, G. L., Reisman, S. J., Iglesias, N., & Gersbach, C. A. (2025) · Molecular Therapy · doi:10.1016/j.ymthe.2025.04.004 · Cited by 43
“28 These innovative strategies highlight the potential for promoters, fusion proteins, and targeted insertions to improve gene therapy efficacy, but this also holds true for other diverse strategies including the integration of regulatory elements within vectors and leveraging cis-and…”
Liu, D., & Kang-Zheng, L. (2025) · Journal of Translational Medicine · doi:10.1186/s12967-025-06524-0 · Cited by 37
“Advanced gene editing technologies like CRISPR/ Cas9, transcription activator-like effector nucleases (TALENs), and zinc finger nucleases (ZFNs), enable precise modifications, improving both the safety and efficacy of immune cell therapies [145].”
Hong, C.-H., Sohn, H., Lee, H.-J., Cho, H.-I., & Kim, T. (2017) · Journal of Immunotherapy · doi:10.1097/cji.0000000000000176 · Cited by 34
The Evolution and Future of CAR T Cells for B‐Cell Acute Lymphoblastic Leukemia
Annesley, C., Summers, C. N., Ceppi, F., & Gardner, R. A. (2017) · Clinical Pharmacology & Therapeutics · doi:10.1002/cpt.950 · Cited by 36
“Preclinical evidence evaluating CAR T-cell performance following construct integration into the TCR locus may enhance long-term CAR expression and reduce T-cell exhaustion.(54) Another potential method of enhancing T-cell persistence has been the focus of preclinical work from the NCI.”
MethodWebster, B., Xiong, Y., Hu, P., Wu, D., Alabanza, L. M., Orentas, R. J., Dropulić, B., & Schneider, D. (2021) · Molecular Therapy · doi:10.1016/j.ymthe.2021.05.006 · Cited by 37
Drag-and-drop genome insertion without DNA cleavage with CRISPR-directed integrasesPreprint
Ioannidi, E. I., Yarnall, M. T. N., Schmitt-Ulms, C., Krajeski, R. N., Lim, J., Villiger, L., Zhou, W., Jiang, K., Roberts, N., Zhang, L., et al. (2021) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2021.11.01.466786 · Cited by 35
CAR‐T‐cell products in solid tumors: Progress, challenges, and strategies
Qian, K., Li, G., Zhang, S., Fu, W., Li, T., Zhao, J., Lei, C., Wang, Y., & Hu, S. (2024) · Interdisciplinary medicine · doi:10.1002/inmd.20230047 · Cited by 36
Optimizing cancer treatment: the synergistic potential of CAR-T cell therapy and CRISPR/Cas9
Amiri, M., Moaveni, A. K., Zolbin, M. M., Shademan, B., & Nourazarian, A. (2024) · Frontiers in Immunology · doi:10.3389/fimmu.2024.1462697 · Cited by 37
“Additionally, CRISPR/Cas9 provides a robust alternative to conventional lentiviral insertion of CARs by avoiding random genome integration and uncontrolled construct expression (107).”
Recent advances in universal chimeric antigen receptor T cell therapy
Jiang, N., Yang, Z., Miao, H., Xing, S., Wang, S., & Li, N. (2025) · Journal of Hematology & Oncology · doi:10.1186/s13045-025-01737-8 · Cited by 41
“This can be achieved by combining gene-editing nucleases with a DNA repair template delivered by a recombinant adeno-associated virus 6 (rAAV6) vector, facilitating precise integration at the desired genomic site[17, 24].”
Perspectives on Manufacturing of High-Quality Cell Therapies
Rivière, I., & Roy, K. (2017) · Molecular Therapy · doi:10.1016/j.ymthe.2017.04.010 · Cited by 34
CRISPR-Cas based targeting of host and viral genes as an antiviral strategy
Koujah, L., Shukla, D., & Naqvi, A. R. (2019) · Seminars in Cell and Developmental Biology · doi:10.1016/j.semcdb.2019.04.004 · Cited by 34
“employed a different approach of targeted delivery of CD19-specific CAR to the T-cell receptor α constant (TRAC) locus using CRISPR/Cas system [146].”
MethodKozani, P. S., Kozani, P. S., & Rahbarizadeh, F. (2021) · Frontiers in Immunology · doi:10.3389/fimmu.2021.765097 · Cited by 37
Engineering CAR T cells for enhanced efficacy and safety
Wu, Y., Huang, Z., Harrison, R. E. S., Liu, L., Zhu, L., Situ, Y., & Wang, Y. (2022) · APL Bioengineering · doi:10.1063/5.0073746 · Cited by 34
Robust T cell activation requires an eIF3-driven burst in T cell receptor translation
Silva, D. D., Ferguson, L., Chin, G. H., Smith, B. E., Apathy, R., Roth, T. L., Blaeschke, F., Kudla, M., Marson, A., Ingolia, N. T., et al. (2021) · eLife · doi:10.7554/elife.74272 · Cited by 32
“Our results using nanoluciferase reporters indicated that eIF3-responsive mRNA 3’-UTR elements could be used to improve chimeric antigen receptor expression and CAR T cell responsiveness (Eyquem et al., 2017; Globerson Levin et al., 2021; Watanabe et al., 2018).”
ResultLiao, C., Wang, Y., Huang, Y., Duan, Y., Liang, Y., Chen, J., Jiang, J., Shang, K., Zhou, C., Gu, Y., et al. (2023) · Advanced Science · doi:10.1002/advs.202207394 · Cited by 33
Alidadi, M., Barzgar, H., Zaman, M., Paevskaya, O. A., Metanat, Y., Khodabandehloo, E., & Moradi, V. (2024) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2024.1491282 · Cited by 36
Central control of dynamic gene circuits governs T cell rest and activation
Arce, M. M., Umhoefer, J. M., Arang, N., Kasinathan, S., Freimer, J. W., Steinhart, Z., Shen, H., Pham, M. T. N., Ota, M., Wadhera, A., et al. (2024) · Nature · doi:10.1038/s41586-024-08314-y · Cited by 38
“Multiple studies have suggested that cycles between T cell rest and activation can influence the durability of adoptive T cell therapies 39,40 .”
Raigani, M., Eftekhari, Z., Adeli, A., & Kazemi‐Lomedasht, F. (2025) · Molecular Therapy — Nucleic Acids · doi:10.1016/j.omtn.2025.102666 · Cited by 36
“140,141 Thus, CRISPR technology has become an essential tool in advancing T cell-based cancer immunotherapies.”
Eyles, J. E., Vessillier, S., Jones, A., Stacey, G., Schneider, C. K., & Price, J. (2018) · Journal of Chemical Technology & Biotechnology · doi:10.1002/jctb.5829 · Cited by 34
Pesch, T., Bonati, L., Kelton, W., Parola, C., Ehling, R. A., Csepregi, L., Kitamura, D., & Reddy, S. T. (2019) · Frontiers in Immunology · doi:10.3389/fimmu.2019.02630 · Cited by 33
Future of CAR T cells in multiple myeloma
Wudhikarn, K., Mailankody, S., & Smith, E. L. (2020) · Hematology · doi:10.1182/hematology.2020000111 · Cited by 35
Parking CAR T Cells in Tumours: Oncolytic Viruses as Valets or Vandals?
Evgin, L., & Vile, R. G. (2021) · Cancers · doi:10.3390/cancers13051106 · Cited by 34
“Retro- or lentiviral vectors are the primary means of introducing stable expression of the CAR into T cells, but random integration into the genome has the potential to lead to insertional mutagenesis and variegated expression of the CAR, thus prompting use of targeted means to introduce the CAR into genomic safe harb…”
Fujiwara, K., Kitaura, M., Tsunei, A., Kusabuka, H., Ogaki, E., & Okada, N. (2021) · International Journal of Molecular Sciences · doi:10.3390/ijms22052476 · Cited by 33
“The expression level of CARs is expected to vary depending not only on the CAR structure but also on the method of CAR gene introduction [40].”
MethodNakamura, K., Yagyu, S., Hirota, S., Tomida, A., Kondo, M., Shigeura, T., Hasegawa, A., Tanaka, M., & Nakazawa, Y. (2021) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2021.03.011 · Cited by 34
Zhu, X., Hu, H., Xiao, Y., Li, Q., Zhong, Z., Yang, J., Zou, P., Cao, Y., Meng, F., Li, W., et al. (2022) · Cancer Letters · doi:10.1016/j.canlet.2022.215668 · Cited by 33
Progresses, Challenges, and Prospects of CRISPR/Cas9 Gene-Editing in Glioma Studies
Kang, X., Wang, Y., Liu, P., Huang, B., Zhou, B., Lu, S., Geng, W., & Tang, H. (2023) · Cancers · doi:10.3390/cancers15020396 · Cited by 37
Guo, T., Ma, D., & Lu, T. K. (2022) · ACS Synthetic Biology · doi:10.1021/acssynbio.1c00236 · Cited by 33
Shakirova, A. I., Karpov, T. E., Komarova, Y., & Lepik, K. V. (2023) · Frontiers in Genome Editing · doi:10.3389/fgeed.2023.1068637 · Cited by 33
“Such activation can be induced during the prolonged culture phase before transduction, which can take from 1 to 2 days for HSPCs and T-cell to 7–14 days for NK cells (Eyquem et al., 2017; De Ravin et al., 2021; Kararoudi, 2021; Clara et al., 2022) or in combination with small molecule expansion stimulators for HSPCs l…”
BackgroundCAR-T cell immunotherapy for ovarian cancer: hushing the silent killer
Nasiri, F., Farrokhi, K., Kozani, P. S., Kancha, M. M., Shokoohi, S. D., & Kozani, P. S. (2023) · Frontiers in Immunology · doi:10.3389/fimmu.2023.1302307 · Cited by 32
“To address this, researchers have used genetic manipulation methods to knock out the TCR a or b chain which renders the produced T-cell product devoid of TCR surface presentation resulting in a signi fi cantly lower rate of alloreactivity (167 – 171).”
MethodTargeting HLA-E-overexpressing cancers with a NKG2A/C switch receptor
Sætersmoen, M., Kotchetkov, I. S., Torralba‐Raga, L., Mansilla‐Soto, J., Sohlberg, E., Krokeide, S. Z., Hammer, Q., Sadelain, M., & Malmberg, K. (2024) · Med · doi:10.1016/j.medj.2024.09.010 · Cited by 33
Self-citationCurrent Non-Viral-Based Strategies to Manufacture CAR-T Cells
Gehrke, L., Gonçalves, V. D. R., Andrae, D., Raskó, T., Ho, P., Einsele, H., Hudecek, M., & Friedel, S. R. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms252413685 · Cited by 34
“Linking CAR expression to TCR signaling via site-specific integration of CARs into the human TRAC locus has been associated with activation-induced downregulation of CAR expression, improved therapy outcome, and reduced exhaustion of T cells after antigen encounter [105].”
Current development of chimeric antigen receptor T-cell therapy
Wang, J., Hu, Y., & Huang, H. (2018) · Stem Cell Investigation · doi:10.21037/sci.2018.11.05 · Cited by 33
“used CRISPR/Cas9 system to direct CAR gene to the T-cell receptor α constant (TRAC) locus, which vastly enhanced CAR T-cell potency (13).”
MethodParola, C., Neumeier, D., Friedensohn, S., Csepregi, L., Tacchio, M. D., Mason, D. M., & Reddy, S. T. (2019) · mAbs · doi:10.1080/19420862.2019.1662691 · Cited by 33
“One of the most noteworthy examples is the use of recombinant adeno-associated viral vectors (rAAV) as HDR donors for therapeutic cell engineering, where targeted knock-in efficiencies up to 40%have beenobserved.(24,25) The field of directed evolution and protein engineering could also benefit from highthroughput geno…”
BackgroundCurrent Perspectives on the Use of off the Shelf CAR-T/NK Cells for the Treatment of Cancer
Cutmore, L. C., & Marshall, J. F. (2021) · Cancers · doi:10.3390/cancers13081926 · Cited by 35
“This simultaneously disrupts the αβ TCR and promotes CAR expression, meaning there are no CAR T cells produced that express the TCR [108,109].”
BackgroundSmirnov, S., Petukhov, A., Levchuk, K., Kulemzin, S. V., Staliarova, A., Lepik, K. V., Shuvalov, O. Y., Zaritskey, A. Y., Daks, A. A., & Fedorova, O. (2021) · Frontiers in Immunology · doi:10.3389/fimmu.2021.780145 · Cited by 34
“The CRISPR/Cas9 efficiency of TCR KO was estimated to be 70% by Eyquem and colleagues (46).”
BackgroundChallenges of CRISPR-Based Gene Editing in Primary T Cells
Rezalotfi, A., Fritz, L. V., Förster, R., & Bošnjak, B. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms23031689 · Cited by 33
Taking T-Cell Oncotherapy Off-the-Shelf
Mo, F., Mamonkin, M., Brenner, M. K., & Heslop, H. E. (2021) · Trends in Immunology · doi:10.1016/j.it.2021.01.004 · Cited by 31
“In this system, the CD19 CAR transgene replaces the endogenous TRAC gene, thus preventing assembly of the full TCR complex and allowing more physiologically regulated CAR expression[40].”
BackgroundCytokine signaling in chimeric antigen receptor T-cell therapy
Kagoya, Y. (2023) · International Immunology · doi:10.1093/intimm/dxad033 · Cited by 31
Unlocking the potential of Tregs: innovations in CAR technology
Cier, C. J. R., Valentini, N., & Lamarche, C. (2023) · Frontiers in Molecular Biosciences · doi:10.3389/fmolb.2023.1267762 · Cited by 33
DNA on the move: mechanisms, functions and applications of transposable elements
Schmitz, M., & Querques, I. (2023) · FEBS Open Bio · doi:10.1002/2211-5463.13743 · Cited by 41
“This is particularly concerning as regulated expression of therapeutic genes in specific genomic contexts is key to the success of gene therapy treatments [39].”
BackgroundRamos, A., Koch, C. E., Liu-Lupo, Y., Hellinger, R. D., Kyung, T., Abbott, K. L., Fröse, J., Goulet, D. R., Gordon, K. S., Eidell, K. P., et al. (2023) · Nature Communications · doi:10.1038/s41467-023-43790-2 · Cited by 32
“In vitro killing assay 750 In vitro CAR-T killing assays were performed using standard methods.(75,76) Briefly, target cells are”
Xiao, K., Lai, Y., Yuan, W., Li, S., Liu, X., Xiao, Z., & Xiao, H. (2023) · Interdisciplinary medicine · doi:10.1002/inmd.20230036 · Cited by 36
Unlocking T cell exhaustion: Insights and implications for CAR-T cell therapy
Xiong, D., Yu, H., & Sun, Z. (2024) · Acta Pharmaceutica Sinica B · doi:10.1016/j.apsb.2024.04.022 · Cited by 31
“A seminal example involves the use of CRISPR/Cas9 to concurrently knock out the endogenous TCR alpha constant (TRAC) locus and knock in a CD19-targeted CAR construct 71 .”
MethodAdvances in cell therapy: progress and challenges in hematological and solid tumors
D'Avanzo, C., Blaeschke, F., Lysandrou, M., Ingelfinger, F., & Zeiser, R. (2024) · Trends in Pharmacological Sciences · doi:10.1016/j.tips.2024.10.016 · Cited by 32
Epigenetic regulation of T cell exhaustion in cancer
Kang, T. G., Johnson, J. T., Zebley, C. C., & Youngblood, B. (2025) · Nature reviews. Cancer · doi:10.1038/s41568-025-00883-y · Cited by 36
“Another study has revealed that CAR T cells preconditioned with IL-7 and IL-15 showed increased ICB responsiveness that was coupled to a preservation of stem-like T cell populations 138 .”
Illuminating the genome-wide activity of genome editors for safe and effective therapeutics
Cheng, Y., & Tsai, S. Q. (2018) · Genome biology · doi:10.1186/s13059-018-1610-2 · Cited by 32
Zeng, J., Tang, S. Y., & Wang, S. (2019) · PLoS ONE · doi:10.1371/journal.pone.0216815 · Cited by 31
Development of CAR-T cells for long-term eradication and surveillance of HIV-1 reservoir
Liu, B., Zhang, W., & Zhang, H. (2019) · Current Opinion in Virology · doi:10.1016/j.coviro.2019.04.004 · Cited by 30
Hsi, P. T., Christianson, R. J., Dubay, R., Lissandrello, C. A., Fiering, J. O., Balestrini, J. L., & Tandon, V. (2019) · Lab on a Chip · doi:10.1039/c9lc00458k · Cited by 31
Controlling Cytokine Release Syndrome to Harness the Full Potential of CAR-Based Cellular Therapy
Thakar, M. S., Kearl, T. J., & Malarkannan, S. (2020) · Frontiers in Oncology · doi:10.3389/fonc.2019.01529 · Cited by 32
“This decreased tonic activation may also decrease CRS (91).”
BackgroundLayer-by-Layer technique as a versatile tool for gene delivery applications
Linnik, D., Tarakanchikova, Y. V., Zyuzin, M. V., Lepik, K. V., Aerts, J. L., Sukhorukov, G. B., & Timin, A. S. (2021) · Expert Opinion on Drug Delivery · doi:10.1080/17425247.2021.1879790 · Cited by 33
“The use of non-viral CAR-T-cell production processes, based either on Sleeping Beauty transposon system [149] or gene editing [150], represents the next step in the development of technology, allowing to dispose of costly and less safe and less efficient retro- and lentiviral vectors.”
MethodJing, R., Jiao, P., Chen, J., Meng, X., Wu, X., Duan, Y., Shang, K., Qian, L., Huang, Y., Liu, J., et al. (2021) · Small Methods · doi:10.1002/smtd.202100071 · Cited by 32
Self-citationNanobody-armed T cells endow CAR-T cells with cytotoxicity against lymphoma cells
Wang, H., Wang, L., Li, Y., Li, G., Zhang, X., Jiang, D., Zhang, Y.-T. Y., Liu, L., Chu, Y., & Xu, G. (2021) · Cancer Cell International · doi:10.1186/s12935-021-02151-z · Cited by 31
“Recent studies also showed that knockout of PD-1 by CRISPR/Cas9 improved the efficiency of CAR-T cells [64].”
BackgroundClara, J., Levy, E. R., Reger, R. N., Barisic, S., Chen, L., Cherkasova, E. A., Chakraborty, M., Allan, D., & Childs, R. W. (2022) · Journal for ImmunoTherapy of Cancer · doi:10.1136/jitc-2021-003804 · Cited by 31
Four challenges to CAR T cells breaking the glass ceiling
Bailey, S. R., Berger, T. R., Graham, C., Larson, R. C., & Maus, M. V. (2022) · European Journal of Immunology · doi:10.1002/eji.202250039 · Cited by 34
Development of HIV-Resistant CAR T Cells by CRISPR/Cas-Mediated CAR Integration into the CCR5 Locus
Rothemejer, F. H., Lauritsen, N. P., Juhl, A. K., Schleimann, M. H., König, S., Søgaard, O. S., Bak, R. O., & Tolstrup, M. (2023) · Viruses · doi:10.3390/v15010202 · Cited by 31
“Targeted integration of a transgene by CRISPR/Cas9 can thus cause disruption of the targeted gene while integrating the transgene [30].”
BackgroundMethodImmunogenetic Metabolomics Reveals Key Enzymes That Modulate CAR T-cell Metabolism and Function
Renauer, P. A., Park, J. J.-Y., Bai, M., Acosta, A., Lee, W., Lin, G. H., Zhang, Y., Dai, X., Wang, G., Errami, Y., et al. (2023) · Cancer Immunology Research · doi:10.1158/2326-6066.cir-22-0565 · Cited by 32
“4–1BB (CD137) transactivation domain, and a CD3 zeta signaling domain (7, 46).”
BackgroundYe, L., Lam, S. Z., Yang, L., Suzuki, K., Zou, Y., Lin, Q., Zhang, Y., Clark, P. R., Peng, L., & Chen, S. (2023) · Nature Biomedical Engineering · doi:10.1038/s41551-023-01058-6 · Cited by 34
“CRISPR is one such technology that is being rapidly and broadly applied to immune cell editing 29,30,64 .”
Remote control of cellular immunotherapy
Huang, K., Liu, L., Huang, Y., Wang, Y., Zhou, Y., & Han, G. (2023) · Nature Reviews Bioengineering · doi:10.1038/s44222-023-00042-8 · Cited by 33
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
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
