Exploration of Digestive Diseases

Table 1. Representative CRISPR-based studies in GC.

Target/Area	Function/Pathway	CRISPR tool	Key findings	Reference
METTL1	tRNA methylation & translation	CRISPR-Cas9 knockout/screening	A novel dependency gene promotes GC proliferation and metastasis.	[2]
OTUD5	Deubiquitination & ferroptosis	CRISPR-Cas9 knockout	p53 suppresses OTUD5, promoting GPX4 degradation and ferroptosis.	[3]
HNRNPA2B1	RNA splicing & chemoresistance	CRISPR-Cas9 knockout	Knockout overcomes chemoresistance in GC stem cells.	[4]

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CRISPR: clustered regularly interspaced short palindromic repeats; GC: gastric cancer; METTL1: methyltransferase-like 1.

Declarations

Author contributions

MZ: Writing—original draft. QS: Visualization. QZ: Writing—review & editing. XY: Conceptualization, Writing—original draft, Writing—review & editing, Supervision. All authors have read and agreed to the published version of the manuscript.

Conflicts of interest

The authors declare no conflicts of interest.

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References

Hartgrink HH, Jansen EP, van Grieken NC, van de Velde CJ. Gastric cancer. Lancet. 2009;374:477–90. [DOI] [PubMed] [PMC]

Zeng Z, Zhang X, Jiang CQ, Zhang YG, Wu X, Li J, et al. Identifying novel therapeutic targets in gastric cancer using genome-wide CRISPR-Cas9 screening. Oncogene. 2022;41:2069–78. [DOI] [PubMed]

Zhang J, Tian T, Li X, Xu K, Lu Y, Li X, et al. p53 inhibits OTUD5 transcription to promote GPX4 degradation and induce ferroptosis in gastric cancer. Clin Transl Med. 2025;15:e70271. [DOI] [PubMed] [PMC]

Yu M, Fei B, Chu S. Targeting HNRNPA2B1 to overcome chemotherapy resistance in gastric cancer stem cells: Mechanisms and therapeutic potential. J Biol Chem. 2025;301:108234. [DOI] [PubMed] [PMC]

Uzun S, Özcan Ö, Gök A, Işık A, Bakır S, Günel-Özcan A, et al. A new CRISPR-mediated Apc knockout allele leads to pyloric gland adenoma-like gastric polyps in mice with C57BL/6;FVB/N mixed background. Animal Model Exp Med. 2025;8:922–9. [DOI] [PubMed] [PMC]

Zhang M, Wang J, Yan J, He C, Xu Q, Liu T, et al. Creation of diversified allelic variation in maize using CRISPR/Cas9 gene editing fragment knockout technology. Shandong Agric Sci. 2024;56:1–5. [DOI]

Lu Z, Li Y, Li Y, Mu Q, Wu Z, Zhang L, et al. Diversification types, mechanisms and application progress of CRISPR-Cas systems. J Econ Zool. 2025.

Wang SW, Gao C, Zheng YM, Yi L, Lu JC, Huang XY, et al. Current applications and future perspective of CRISPR/Cas9 gene editing in cancer. Mol Cancer. 2022;21:57. [DOI] [PubMed] [PMC]

Ishino Y, Shinagawa H, Makino K, Amemura M, Nakata A. Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product. J Bacteriol. 1987;169:5429–33. [DOI] [PubMed] [PMC]

10.

Makarova KS, Wolf YI, Alkhnbashi OS, Costa F, Shah SA, Saunders SJ, et al. An updated evolutionary classification of CRISPR-Cas systems. Nat Rev Microbiol. 2015;13:722–36. [DOI] [PubMed] [PMC]

11.

Hille F, Richter H, Wong SP, Bratovič M, Ressel S, Charpentier E. The Biology of CRISPR-Cas: Backward and Forward. Cell. 2018;172:1239–59. [DOI] [PubMed]

12.

Tan L, Ye B, Yin B. Research and application of engineered gRNA technology in CRISPR/Cas system. Chin J Biochem Mol Biol. 2024;40:1078–92. [DOI]

13.

Makarova KS, Wolf YI, Koonin EV. Classification and Nomenclature of CRISPR-Cas Systems: Where from Here? CRISPR J. 2018;1:325–36. [DOI] [PubMed] [PMC]

14.

Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S, et al. CRISPR provides acquired resistance against viruses in prokaryotes. Science. 2007;315:1709–12. [DOI] [PubMed]

15.

Makarova KS, Wolf YI, Iranzo J, Shmakov SA, Alkhnbashi OS, Brouns SJJ, et al. Evolutionary classification of CRISPR-Cas systems: a burst of class 2 and derived variants. Nat Rev Microbiol. 2020;18:67–83. [DOI] [PubMed] [PMC]

16.

Pyzocha NK, Chen S. Diverse Class 2 CRISPR-Cas Effector Proteins for Genome Engineering Applications. ACS Chem Biol. 2018;13:347–56. [DOI] [PubMed] [PMC]

17.

Happi Mbakam C, Lamothe G, Tremblay G, Tremblay JP. CRISPR-Cas9 Gene Therapy for Duchenne Muscular Dystrophy. Neurotherapeutics. 2022;19:931-41. [DOI] [PubMed] [PMC]

18.

Smithies O, Gregg RG, Boggs SS, Koralewski MA, Kucherlapati RS. Insertion of DNA sequences into the human chromosomal beta-globin locus by homologous recombination. Nature. 1985;317:230–4. [DOI] [PubMed]

19.

Thomas KR, Capecchi MR. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell. 1987;51:503–12. [DOI] [PubMed]

20.

Kuehn MR, Bradley A, Robertson EJ, Evans MJ. A potential animal model for Lesch-Nyhan syndrome through introduction of HPRT mutations into mice. Nature. 1987;326:295–8. [DOI] [PubMed]

21.

Zeng S, Lei S, Qu C, Wang Y, Teng S, Huang P. CRISPR/Cas-based gene editing in therapeutic strategies for beta-thalassemia. Hum Genet. 2023;142:1677–703. [DOI] [PubMed]

22.

Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. 2012;337:816–21. [DOI] [PubMed] [PMC]

23.

Su S, Zou Z, Chen F, Ding N, Du J, Shao J, et al. CRISPR-Cas9-mediated disruption of PD-1 on human T cells for adoptive cellular therapies of EBV positive gastric cancer. Oncoimmunology. 2016;6:e1249558. [DOI] [PubMed] [PMC]

24.

Hartenian E, Doench JG. Genetic screens and functional genomics using CRISPR/Cas9 technology. FEBS J. 2015;282:1383–93. [DOI] [PubMed]

25.

Rezaei Z, Ranjbaran J, Safarpour H, Nomiri S, Salmani F, Chamani E, et al. Identification of early diagnostic biomarkers via WGCNA in gastric cancer. Biomed Pharmacother. 2022;145:112477. [DOI] [PubMed]

26.

Bębnowska D, Grywalska E, Niedźwiedzka-Rystwej P, Sosnowska-Pasiarska B, Smok-Kalwat J, Pasiarski M, et al. CAR-T Cell Therapy-An Overview of Targets in Gastric Cancer. J Clin Med. 2020;9:1894. [DOI] [PubMed] [PMC]

27.

Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead EH, et al. Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Cell. 2014;159:647–61. [DOI] [PubMed] [PMC]

28.

Wang Q, Chen C, Ding Q, Zhao Y, Wang Z, Chen J, et al. METTL3-mediated m⁶A modification of HDGF mRNA promotes gastric cancer progression and has prognostic significance. Gut. 2020;69:1193–205. [DOI] [PubMed]

29.

Roux PP, Topisirovic I. Signaling Pathways Involved in the Regulation of mRNA Translation. Mol Cell Biol. 2018;38:e00070–18. [DOI] [PubMed] [PMC]

30.

Xiao S, Li S, Yuan Z, Zhou L. Pyrroline-5-carboxylate reductase 1 (PYCR1) upregulation contributes to gastric cancer progression and indicates poor survival outcome. Ann Transl Med. 2020;8:937. [DOI] [PubMed] [PMC]

31.

Burr ML, Sparbier CE, Chan YC, Williamson JC, Woods K, Beavis PA, et al. CMTM6 maintains the expression of PD-L1 and regulates anti-tumour immunity. Nature. 2017;549:101–5. [DOI] [PubMed] [PMC]

32.

Manguso RT, Pope HW, Zimmer MD, Brown FD, Yates KB, Miller BC, et al. In vivo CRISPR screening identifies Ptpn2 as a cancer immunotherapy target. Nature. 2017;547:413–8. [DOI] [PubMed] [PMC]

33.

Lin H, Cheng J, Mu W, Zhou J, Zhu L. Advances in Universal CAR-T Cell Therapy. Front Immunol. 2021;12:744823. [DOI] [PubMed] [PMC]

34.

Li C, Mei H, Hu Y. Applications and explorations of CRISPR/Cas9 in CAR T-cell therapy. Brief Funct Genomics. 2020;19:175–82. [DOI] [PubMed] [PMC]

35.

Ohaegbulam KC, Assal A, Lazar-Molnar E, Yao Y, Zang X. Human cancer immunotherapy with antibodies to the PD-1 and PD-L1 pathway. Trends Mol Med. 2015;21:24–33. [DOI] [PubMed] [PMC]

36.

Stadtmauer EA, Fraietta JA, Davis MM, Cohen AD, Weber KL, Lancaster E, et al. CRISPR-engineered T cells in patients with refractory cancer. Science. 2020;367:eaba7365. [DOI] [PubMed] [PMC]

37.

Giuffrida L, Sek K, Henderson MA, Lai J, Chen AXY, Meyran D, et al. CRISPR/Cas9 mediated deletion of the adenosine A2A receptor enhances CAR T cell efficacy. Nat Commun. 2021;12:3236. [DOI] [PubMed] [PMC]

38.

Eshhar Z, Waks T, Gross G, Schindler DG. Specific activation and targeting of cytotoxic lymphocytes through chimeric single chains consisting of antibody-binding domains and the gamma or zeta subunits of the immunoglobulin and T-cell receptors. Proc Natl Acad Sci U S A. 1993;90:720–4. [DOI] [PubMed] [PMC]

39.

Finney HM, Lawson AD, Bebbington CR, Weir AN. Chimeric receptors providing both primary and costimulatory signaling in T cells from a single gene product. J Immunol. 1998;161:2791–7. [PubMed]

40.

Davenport AJ, Jenkins MR, Cross RS, Yong CS, Prince HM, Ritchie DS, et al. CAR-T Cells Inflict Sequential Killing of Multiple Tumor Target Cells. Cancer Immunol Res. 2015;3:483–94. [DOI] [PubMed]

41.

Xiong W, Chen Y, Kang X, Chen Z, Zheng P, Hsu YH, et al. Immunological Synapse Predicts Effectiveness of Chimeric Antigen Receptor Cells. Mol Ther. 2021;29:1349–51. [DOI] [PubMed] [PMC]

42.

Panagopoulou TI, Rafiq QA. CAR-T immunotherapies: Biotechnological strategies to improve safety, efficacy and clinical outcome through CAR engineering. Biotechnol Adv. 2019;37:107411. [DOI] [PubMed]

43.

Chmielewski M, Abken H. TRUCKs: the fourth generation of CARs. Expert Opin Biol Ther. 2015;15:1145–54. [DOI] [PubMed]

44.

Hombach AA, Abken H. Costimulation by chimeric antigen receptors revisited the T cell antitumor response benefits from combined CD28-OX40 signalling. Int J Cancer. 2011;129:2935–44. [DOI] [PubMed]

45.

Salter AI, Ivey RG, Kennedy JJ, Voillet V, Rajan A, Alderman EJ, et al. Phosphoproteomic analysis of chimeric antigen receptor signaling reveals kinetic and quantitative differences that affect cell function. Sci Signal. 2018;11:eaat6753. [DOI] [PubMed] [PMC]

46.

Till BG, Jensen MC, Wang J, Qian X, Gopal AK, Maloney DG, et al. CD20-specific adoptive immunotherapy for lymphoma using a chimeric antigen receptor with both CD28 and 4-1BB domains: pilot clinical trial results. Blood. 2012;119:3940–50. [DOI] [PubMed] [PMC]

47.

Ramello MC, Benzaïd I, Kuenzi BM, Lienlaf-Moreno M, Kandell WM, Santiago DN, et al. An immunoproteomic approach to characterize the CAR interactome and signalosome. Sci Signal. 2019;12:eaap9777. [DOI] [PubMed] [PMC]

48.

Qi C, Xie T, Zhou J, Wang X, Gong J, Zhang X, et al. CT041 CAR T cell therapy for Claudin18.2-positive metastatic pancreatic cancer. J Hematol Oncol. 2023;16:102. [DOI] [PubMed] [PMC]

49.

Fleuranvil RF, Rustgi N, James M, Hammer Z, Ryan TJ, Reyes J, et al. Abstract 2942: Efficacy of a claudin 18.2/TGFβ bi-specific CAR-T in preclinical mouse models of gastric cancer. Cancer Res. 2025;85:2942. [DOI]

50.

Shi H, Li A, Dai Z, Xue J, Zhao Q, Tian J, et al. IL-15 armoring enhances the antitumor efficacy of claudin 18.2-targeting CAR-T cells in syngeneic mouse tumor models. Front Immunol. 2023;14:1165404. [DOI]

51.

Dabas P, Danda A. Revolutionizing cancer treatment: a comprehensive review of CAR-T cell therapy. Med Oncol. 2023;40:275. [DOI] [PubMed]

52.

Watanabe N, Mo F, McKenna MK. Impact of Manufacturing Procedures on CAR T Cell Functionality. Front Immunol. 2022;13:876339. [DOI] [PubMed] [PMC]

53.

Uscanga-Palomeque AC, Chávez-Escamilla AK, Alvizo-Báez CA, Saavedra-Alonso S, Terrazas-Armendáriz LD, Tamez-Guerra RS, et al. CAR-T Cell Therapy: From the Shop to Cancer Therapy. Int J Mol Sci. 2023;24:15688. [DOI] [PubMed] [PMC]

54.

Zhao Z, Chen Y, Francisco NM, Zhang Y, Wu M. The application of CAR-T cell therapy in hematological malignancies: advantages and challenges. Acta Pharm Sin B. 2018;8:539–51. [DOI] [PubMed] [PMC]

55.

Ma HY, Das J, Prendergast C, De Jong D, Braumuller B, Paily J, et al. Advances in CAR T Cell Therapy for Non-Small Cell Lung Cancer. Curr Issues Mol Biol. 2023;45:9019–38. [DOI] [PubMed] [PMC]

56.

Pant A, Lim M. CAR-T Therapy in GBM: Current Challenges and Avenues for Improvement. Cancers (Basel). 2023;15:1249. [DOI] [PubMed] [PMC]

57.

Kong Y, Li J, Zhao X, Wu Y, Chen L. CAR-T cell therapy: developments, challenges, and expanded applications from cancer to autoimmunity. Front Immunol. 2025;15:1519671. [DOI] [PubMed] [PMC]

58.

Tao R, Han X, Bai X, Yu J, Ma Y, Chen W, et al. Revolutionizing cancer treatment: enhancing CAR-T cell therapy with CRISPR/Cas9 gene editing technology. Front Immunol. 2024;15:1354825. [DOI] [PubMed] [PMC]

59.

Rautela J, Surgenor E, Huntington ND. Drug target validation in primary human natural killer cells using CRISPR RNP. J Leukoc Biol. 2020;108:1397–408. [DOI] [PubMed]

60.

Baumeister SHC, Mohan GS, Elhaddad A, Lehmann L. Cytokine Release Syndrome and Associated Acute Toxicities in Pediatric Patients Undergoing Immune Effector Cell Therapy or Hematopoietic Cell Transplantation. Front Oncol. 2022;12:841117. [DOI] [PubMed] [PMC]

61.

Kleinstiver BP, Pattanayak V, Prew MS, Tsai SQ, Nguyen NT, Zheng Z, et al. High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature. 2016;529:490–5. [DOI] [PubMed] [PMC]

62.

Nguyen DT, Ogando-Rivas E, Liu R, Wang T, Rubin J, Jin L, et al. CAR T Cell Locomotion in Solid Tumor Microenvironment. Cells. 2022;11:1974. [DOI] [PubMed] [PMC]

63.

Labanieh L, Majzner RG, Klysz D, Sotillo E, Fisher CJ, Vilches-Moure JG, et al. Enhanced safety and efficacy of protease-regulated CAR-T cell receptors. Cell. 2022;185:1745–63.e22. [DOI] [PubMed] [PMC]

64.

Chen C, Ji W, Niu Y. Primate Organoids and Gene-Editing Technologies toward Next-Generation Biomedical Research. Trends Biotechnol. 2021;39:1332–42. [DOI] [PubMed]

65.

Tuveson D, Clevers H. Cancer modeling meets human organoid technology. Science. 2019;364:952–5. [DOI] [PubMed]

66.

Lo YH, Kolahi KS, Du Y, Chang CY, Krokhotin A, Nair A, et al. A CRISPR/Cas9-Engineered ARID1A-Deficient Human Gastric Cancer Organoid Model Reveals Essential and Nonessential Modes of Oncogenic Transformation. Cancer Discov. 2021;11:1562–81. [DOI] [PubMed] [PMC]

67.

Nanki K, Toshimitsu K, Takano A, Fujii M, Shimokawa M, Ohta Y, et al. Divergent Routes toward Wnt and R-spondin Niche Independency during Human Gastric Carcinogenesis. Cell. 2018;174:856–69.e17. [DOI] [PubMed]

68.

Zhao Y, Li S, Zhu L, Huang M, Xie Y, Song X, et al. Personalized drug screening using patient-derived organoid and its clinical relevance in gastric cancer. Cell Rep Med. 2024;5:101627. [DOI] [PubMed] [PMC]

69.

Bulcha JT, Wang Y, Ma H, Tai PWL, Gao G. Viral vector platforms within the gene therapy landscape. Signal Transduct Target Ther. 2021;6:53. [DOI] [PubMed] [PMC]

70.

Zhang S, Shen J, Li D, Cheng Y. Strategies in the delivery of Cas9 ribonucleoprotein for CRISPR/Cas9 genome editing. Theranostics. 2021;11:614–48. [DOI] [PubMed] [PMC]

71.

Fajrial AK, He QQ, Wirusanti NI, Slansky JE, Ding X. A review of emerging physical transfection methods for CRISPR/Cas9-mediated gene editing. Theranostics. 2020;10:5532–49. [DOI] [PubMed] [PMC]

72.

Behr M, Zhou J, Xu B, Zhang H. In vivo delivery of CRISPR-Cas9 therapeutics: Progress and challenges. Acta Pharm Sin B. 2021;11:2150–71. [DOI] [PubMed] [PMC]

73.

Zhao R, Meng L, Feng J, Liu P, Jia T, Zeng W, et al. Optimization of CRISPR/Cas9 system for plasmid electroporation delivery to Jurkat cells. Chin J Cell Biol. 2019;41:702–9. [DOI]

74.

Wei T, Cheng Q, Farbiak L, Anderson DG, Langer R, Siegwart DJ. Delivery of Tissue-Targeted Scalpels: Opportunities and Challenges for In Vivo CRISPR/Cas-Based Genome Editing. ACS Nano. 2020;14:9243–62. [DOI] [PubMed] [PMC]

75.

Lei E, Miao M, Cao Y, Yang J. Application of CRISPR/Cas9 gene editing technology in the study of viral infectious diseases. Mil Med. 2021;45:384–9. [DOI]

76.

Adashi EY, Cohen IG. Heritable Human Genome Editing: The International Commission Report. JAMA. 2020;324:1941–2. [DOI] [PubMed]

77.

Turocy J, Adashi EY, Egli D. Heritable human genome editing: Research progress, ethical considerations, and hurdles to clinical practice. Cell. 2021;184:1561–74. [DOI] [PubMed]

78.

Tsuchida CA, Brandes N, Bueno R, Trinidad M, Mazumder T, Yu B, et al. Mitigation of chromosome loss in clinical CRISPR-Cas9-engineered T cells. Cell. 2023;186:4567–82.e20. [DOI] [PubMed] [PMC]

79.

Imbach K, Patel A, Levine AD. Ethical Considerations in the Translation of CAR-T Cell Therapies. Cell Gene Ther Insights. 2018;4:295–307. [DOI]