nav emailalert searchbtn searchbox tablepage yinyongbenwen piczone journalimg journalInfo journalinfonormal searchdiv searchzone qikanlogo popupnotification paper paperNew
CRISPR-Cas系统的多样化类型、机制及其应用进展
基金项目(Foundation): 内蒙古自治区科技计划项目(2023YFHH0076); 国家自然科学基金项目(32160772); 内蒙古自治区自然科学基金项目(2024ZD14); 内蒙古自治区高等学校创新团队发展计划项目(NMGIRT2322)
邮箱(Email): jsliu588@163.com;13947196432@163.com
DOI:
发布时间: 2025-04-16
出版时间: 2025-04-16
网络发布时间: 2025-04-16
移动端阅读
摘要:

过去,进化是由随机的基因突变和自然选择驱动的。现在,CRISPR (Clustered Regularly-Interspaced Short Palindromic Repeats)-Cas (CRISPR-associated proteins)可以自由改变生命方向,成为新型的基因编辑免疫系统分子工具。由于CRISPR-Cas系统精确、简单、高效、成本低并可与多种技术结合等优点,在基因功能研究、农牧业基因编辑育种、人类医学疾病治疗、药物研发和环境治理等领域都具有极大的应用前景。根据CRISPR序列结构特征和Cas蛋白的差异,在基因编辑领域主要存在2种类别6个类型的CRISPR,多种Cas蛋白及其辅助性蛋白,目前应用范围最广的基因编辑工具是CRISPR-Cas3、CRISPR-Cas9和CRISPR-Cas12系统。在这篇综述中,我们对CRISPR-Cas的不同类型、不同Cas蛋白的功能机制、现存问题以及应用挑战和前景进行讨论,阐明这一系统的多样性和复杂性,为未来几十年Cas蛋白在基因编辑、疾病防治等科学研究的使用提供理论参考。

Abstract:

In the past, evolution was driven by random gene mutations and natural selection. Now, CRISPR (Clustered Regularly-Interspaced Short Palindromic Repeats)-Cas (CRISPR-associated proteins) can freely alter the direction of life and become a new type of gene editing immune system molecular tool. Due to the advantages of precision, simplicity, high efficiency, low cost, and compatibility with various technologies, CRISPR-Cas systems have great application prospects in the fields of gene function research, agricultural and animal husbandry gene editing breeding, human medical disease treatment, drug development, and environmental governance. According to the structural features of CRISPR sequences and the differences in Cas proteins, there are mainly two categories and six types of CRISPR in gene editing, as well as various Cas proteins and auxiliary proteins. The most widely used gene editing tools at present are the CRISPR-Cas3, CRISPR-Cas9, and CRISPR-Cas12 systems. In this review, we discuss the different types, functions and mechanisms of different Cas proteins, existing problems, and application challenges and prospects of the CRISPR-Cas system, to clarify the diversity and complexity of the system, and provide theoretical reference for the use of Cas proteins in scientific research such as gene editing and disease prevention in the coming decades.

参考文献

1. 张茂林; 王娟; 严佳丽; 何春梅; 徐倩; 刘铁山; 董瑞; 刘春晓; 关海英; 刘强; 等. 利用CRISPR/Cas9基因编辑片段敲除技术创制玉米多样化等位变异. 山东农业科学. 2024, 56, 1-5, doi:10.14083/j.issn.1001-4942.2024.09.001.

2. Xu, L.; Wang, J.; Liu, Y.; Xie, L.; Su, B.; Mou, D.; Wang, L.; Liu, T.; Wang, X.; Zhang, B.; et al. CRISPR-Edited Stem Cells in a Patient with HIV and Acute Lymphocytic Leukemia. The New England journal of medicine 2019, 381, 1240-1247, doi:10.1056/NEJMoa1817426.

3. Yue, Y.; Xu, W.; Kan, Y.; Zhao, H.Y.; Zhou, Y.; Song, X.; Wu, J.; Xiong, J.; Goswami, D.; Yang, M.; et al. Extensive germline genome engineering in pigs. Nature biomedical engineering 2021, 5, 134-143, doi:10.1038/s41551-020-00613-9.

4. 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. Journal of bacteriology 1987, 169, 5429-5433, doi:10.1128/jb.169.12.5429-5433.1987.

5. Bolotin, A.; Quinquis, B.; Sorokin, A.; Ehrlich, S.D. Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin. Microbiology (Reading, England) 2005, 151, 2551-2561, doi:10.1099/mic.0.28048-0.

6. Jore, M.M.; Lundgren, M.; van Duijn, E.; Bultema, J.B.; Westra, E.R.; Waghmare, S.P.; Wiedenheft, B.; Pul, U.; Wurm, R.; Wagner, R.; et al. Structural basis for CRISPR RNA-guided DNA recognition by Cascade. Nature structural & molecular biology 2011, 18, 529-536, doi:10.1038/nsmb.2019.

7. Charpentier, E.; Doudna, J.A. Biotechnology: Rewriting a genome. Nature 2013, 495, 50-51, doi:10.1038/495050a.

8. Cong, L.; Ran, F.A.; Cox, D.; Lin, S.; Barretto, R.; Habib, N.; Hsu, P.D.; Wu, X.; Jiang, W.; Marraffini, L.A.; et al. Multiplex genome engineering using CRISPR/Cas systems. Science (New York, N.Y.) 2013, 339, 819-823, doi:10.1126/science.1231143.

9. Lee, J.; Mou, H.; Ibraheim, R.; Liang, S.Q.; Liu, P.; Xue, W.; Sontheimer, E.J. Tissue-restricted genome editing in vivo specified by microRNA-repressible anti-CRISPR proteins. RNA (New York, N.Y.) 2019, 25, 1421-1431, doi:10.1261/rna.071704.119.

10. Hwang, S.; Maxwell, K.L. Meet the Anti-CRISPRs: Widespread Protein Inhibitors of CRISPR-Cas Systems. The CRISPR journal 2019, 2, 23-30, doi:10.1089/crispr.2018.0052.

11. Stanley, S.Y.; Maxwell, K.L. Phage-Encoded Anti-CRISPR Defenses. Annual review of genetics 2018, 52, 445-464, doi:10.1146/annurev-genet-120417-031321.

12. Koonin, E.V.; Makarova, K.S. Origins and evolution of CRISPR-Cas systems. Philosophical transactions of the Royal Society of London. Series B, Biological sciences 2019, 374, 20180087, doi:10.1098/rstb.2018.0087.

13. Hille, F.; Richter, H.; Wong, S.P.; Bratovi?, M.; Ressel, S.; Charpentier, E. The Biology of CRISPR-Cas: Backward and Forward. Cell 2018, 172, 1239-1259, doi:10.1016/j.cell.2017.11.032.

14. 谈鎏; 叶邦策; 尹斌成. CRISPR/Cas系统中工程化gRNA技术的研究和应用. 中国生物化学与分子生物学报. 2024, 40, 1078-1092, doi:10.13865/j.cnki.cjbmb.2024.06.1027.

15. Makarova, K.S.; Wolf, Y.I.; Koonin, E.V. Classification and Nomenclature of CRISPR-Cas Systems: Where from Here? The CRISPR journal 2018, 1, 325-336, doi:10.1089/crispr.2018.0033.

16. 赵雅楠. RRM识别癌相关microRNA的分子机制研究. 硕士, 2022.

17. 张钰雯; 俞晨霖; 戴心忱; 肖易倍; 陆美玲. Ⅰ型CRISPR-Cas系统效应物的结构特征及其在基因编辑领域的应用. 中国药科大学学报. 2021, 52, 675-683.

18. 徐朋; 冯书营; 李姝璇; 赵称心; 张博雅; 刘晓. CRISPR/Cas系统在微藻基因编辑研究中的进展. 郑州大学学报(医学版). 2024, 59, 629-634, doi:10.13705/j.issn.1671-6825.2023.11.012.

19. Coleman, G.A.; Davín, A.A.; Mahendrarajah, T.A.; Szánthó, L.L.; Spang, A.; Hugenholtz, P.; Sz?ll?si, G.J.; Williams, T.A. A rooted phylogeny resolves early bacterial evolution. Science (New York, N.Y.) 2021, 372, doi:10.1126/science.abe0511.

20. Molina, R.; Sofos, N.; Montoya, G. Structural basis of CRISPR-Cas Type III prokaryotic defence systems. Current opinion in structural biology 2020, 65, 119-129, doi:10.1016/j.sbi.2020.06.010.

21. 洪甜. CRISPR系统Cas7-11效应蛋白的结构和功能研究. 硕士, 2023.

22. 薛振勇. 利用荧光技术研究DinG和RecD2解旋酶与DNA的相互作用. 硕士, 2020.

23. Shmakov, S.; Smargon, A.; Scott, D.; Cox, D.; Pyzocha, N.; Yan, W.; Abudayyeh, O.O.; Gootenberg, J.S.; Makarova, K.S.; Wolf, Y.I.; et al. Diversity and evolution of class 2 CRISPR-Cas systems. Nature reviews. Microbiology 2017, 15, 169-182, doi:10.1038/nrmicro.2016.184.

24. 周建; 任雪梅; 王馨; 李卓. CRISPR/Cas核酸检测系统的研究进展和未来挑战. 检验医学. 2024, 39, 608-614.

25. 魏家玮. 结核分枝杆菌Cas1和RNA结合蛋白在结核致病及免疫逃逸中的作用. 博士, 2020.

26. Koonin, E.V. CRISPR: a new principle of genome engineering linked to conceptual shifts in evolutionary biology. Biology & philosophy 2019, 34, 9, doi:10.1007/s10539-018-9658-7.

27. Mahas, A.; Mahfouz, M. Engineering virus resistance via CRISPR-Cas systems. Current opinion in virology 2018, 32, 1-8, doi:10.1016/j.coviro.2018.06.002.

28. Murugan, K.; Babu, K.; Sundaresan, R.; Rajan, R.; Sashital, D.G. The Revolution Continues: Newly Discovered Systems Expand the CRISPR-Cas Toolkit. Molecular cell 2017, 68, 15-25, doi:10.1016/j.molcel.2017.09.007.

29. Watters, K.E.; Fellmann, C.; Bai, H.B.; Ren, S.M.; Doudna, J.A. Systematic discovery of natural CRISPR-Cas12a inhibitors. Science (New York, N.Y.) 2018, 362, 236-239, doi:10.1126/science.aau5138.

30. Hillary, V.E.; Ceasar, S.A. A Review on the Mechanism and Applications of CRISPR/Cas9/Cas12/Cas13/Cas14 Proteins Utilized for Genome Engineering. Molecular biotechnology 2023, 65, 311-325, doi:10.1007/s12033-022-00567-0.

31. Abudayyeh, O.O.; Gootenberg, J.S.; Essletzbichler, P.; Han, S.; Joung, J.; Belanto, J.J.; Verdine, V.; Cox, D.B.T.; Kellner, M.J.; Regev, A.; et al. RNA targeting with CRISPR-Cas13. Nature 2017, 550, 280-284, doi:10.1038/nature24049.

32. O'Connell, M.R. Molecular Mechanisms of RNA Targeting by Cas13-containing Type VI CRISPR-Cas Systems. Journal of molecular biology 2019, 431, 66-87, doi:10.1016/j.jmb.2018.06.029.

33. Smargon, A.A.; Cox, D.B.T.; Pyzocha, N.K.; Zheng, K.; Slaymaker, I.M.; Gootenberg, J.S.; Abudayyeh, O.A.; Essletzbichler, P.; Shmakov, S.; Makarova, K.S.; et al. Cas13b Is a Type VI-B CRISPR-Associated RNA-Guided RNase Differentially Regulated by Accessory Proteins Csx27 and Csx28. Molecular cell 2017, 65, 618-630.e617, doi:10.1016/j.molcel.2016.12.023.

34. Abudayyeh, O.O.; Gootenberg, J.S.; Konermann, S.; Joung, J.; Slaymaker, I.M.; Cox, D.B.; Shmakov, S.; Makarova, K.S.; Semenova, E.; Minakhin, L.; et al. C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science (New York, N.Y.) 2016, 353, aaf5573, doi:10.1126/science.aaf5573.

35. Samai, P.; Pyenson, N.; Jiang, W.; Goldberg, G.W.; Hatoum-Aslan, A.; Marraffini, L.A. Co-transcriptional DNA and RNA Cleavage during Type III CRISPR-Cas Immunity. Cell 2015, 161, 1164-1174, doi:10.1016/j.cell.2015.04.027.

36. Makarova, K.S.; Wolf, Y.I.; Alkhnbashi, O.S.; Costa, F.; Shah, S.A.; Saunders, S.J.; Barrangou, R.; Brouns, S.J.; Charpentier, E.; Haft, D.H.; et al. An updated evolutionary classification of CRISPR-Cas systems. Nature reviews. Microbiology 2015, 13, 722-736, doi:10.1038/nrmicro3569.

37. 吴祉乐. 检测已知CRISPR-Cas系统及发掘未知Cas蛋白. 硕士, 2021.

38. 毛旭丹. 类组蛋白StpA对大肠杆菌I-E型CRISPR-Cas系统调控机制研究. 硕士, 2020.

39. 刘茜. 细菌Ⅲ-E型CRISPR-Cas免疫系统的结构和分子机制研究. 博士, 2023.

40. 李振国. 结核分枝杆菌Cas6蛋白与巨噬细胞内蛋白相互作用的初步探究. 硕士, 2017.

41. Makarova, K.S.; Haft, D.H.; Barrangou, R.; Brouns, S.J.; Charpentier, E.; Horvath, P.; Moineau, S.; Mojica, F.J.; Wolf, Y.I.; Yakunin, A.F.; et al. Evolution and classification of the CRISPR-Cas systems. Nature reviews. Microbiology 2011, 9, 467-477, doi:10.1038/nrmicro2577.

42. Ali, Z.; Mahas, A.; Mahfouz, M. CRISPR/Cas13 as a Tool for RNA Interference. Trends in plant science 2018, 23, 374-378, doi:10.1016/j.tplants.2018.03.003.

43. Almendros, C.; Nobrega, F.L.; McKenzie, R.E.; Brouns, S.J.J. Cas4-Cas1 fusions drive efficient PAM selection and control CRISPR adaptation. Nucleic acids research 2019, 47, 5223-5230, doi:10.1093/nar/gkz217.

44. Nasef, M.; Muffly, M.C.; Beckman, A.B.; Rowe, S.J.; Walker, F.C.; Hatoum-Aslan, A.; Dunkle, J.A. Regulation of cyclic oligoadenylate synthesis by the Staphylococcus epidermidis Cas10-Csm complex. RNA (New York, N.Y.) 2019, 25, 948-962, doi:10.1261/rna.070417.119.

45. Lin, P.; Pu, Q.; Shen, G.; Li, R.; Guo, K.; Zhou, C.; Liang, H.; Jiang, J.; Wu, M. CdpR Inhibits CRISPR-Cas Adaptive Immunity to Lower Anti-viral Defense while Avoiding Self-Reactivity. iScience 2019, 13, 55-68, doi:10.1016/j.isci.2019.02.005.

46. Trasanidou, D.; Gerós, A.S.; Mohanraju, P.; Nieuwenweg, A.C.; Nobrega, F.L.; Staals, R.H.J. Keeping crispr in check: diverse mechanisms of phage-encoded anti-crisprs. FEMS microbiology letters 2019, 366, doi:10.1093/femsle/fnz098.

47. Maxwell, K.L. The Anti-CRISPR Story: A Battle for Survival. Molecular cell 2017, 68, 8-14, doi:10.1016/j.molcel.2017.09.002.

48. Chaudhary, K.; Chattopadhyay, A.; Pratap, D. Anti-CRISPR proteins: Counterattack of phages on bacterial defense (CRISPR/Cas) system. Journal of cellular physiology 2018, 233, 57-59, doi:10.1002/jcp.25877.

49. Landsberger, M.; Gandon, S.; Meaden, S.; Rollie, C.; Chevallereau, A.; Chabas, H.; Buckling, A.; Westra, E.R.; van Houte, S. Anti-CRISPR Phages Cooperate to Overcome CRISPR-Cas Immunity. Cell 2018, 174, 908-916.e912, doi:10.1016/j.cell.2018.05.058.

50. Pawluk, A.; Davidson, A.R.; Maxwell, K.L. Anti-CRISPR: discovery, mechanism and function. Nature reviews. Microbiology 2018, 16, 12-17, doi:10.1038/nrmicro.2017.120.

51. He, F.; Bhoobalan-Chitty, Y.; Van, L.B.; Kjeldsen, A.L.; Dedola, M.; Makarova, K.S.; Koonin, E.V.; Brodersen, D.E.; Peng, X. Anti-CRISPR proteins encoded by archaeal lytic viruses inhibit subtype I-D immunity. Nature microbiology 2018, 3, 461-469, doi:10.1038/s41564-018-0120-z.

52. Hong, S.; Ka, D.; Yoon, S.J.; Suh, N.; Jeong, M.; Suh, J.Y.; Bae, E. CRISPR RNA and anti-CRISPR protein binding to the Xanthomonas albilineans Csy1-Csy2 heterodimer in the type I-F CRISPR-Cas system. The Journal of biological chemistry 2018, 293, 2744-2754, doi:10.1074/jbc.RA117.001611.

53. Wang, J.; Ma, J.; Cheng, Z.; Meng, X.; You, L.; Wang, M.; Zhang, X.; Wang, Y. A CRISPR evolutionary arms race: structural insights into viral anti-CRISPR/Cas responses. Cell research 2016, 26, 1165-1168, doi:10.1038/cr.2016.103.

54. van Erp, P.B.; Jackson, R.N.; Carter, J.; Golden, S.M.; Bailey, S.; Wiedenheft, B. Mechanism of CRISPR-RNA guided recognition of DNA targets in Escherichia coli. Nucleic acids research 2015, 43, 8381-8391, doi:10.1093/nar/gkv793.

55. 王小飞. Anti-CRISPR蛋白AcrF3抑制I-F型CRISPR-Cas系统的分子机制研究. 博士, 2018.

57. Li, J.; Xu, Z.; Chupalov, A.; Marchisio, M.A. Anti-CRISPR-based biosensors in the yeast S. cerevisiae. Journal of biological engineering 2018, 12, 11, doi:10.1186/s13036-018-0101-z.

58. Kim, I.; Jeong, M.; Ka, D.; Han, M.; Kim, N.K.; Bae, E.; Suh, J.Y. Solution structure and dynamics of anti-CRISPR AcrIIA4, the Cas9 inhibitor. Scientific reports 2018, 8, 3883, doi:10.1038/s41598-018-22177-0.

59. Liu, L.; Yin, M.; Wang, M.; Wang, Y. Phage AcrIIA2 DNA Mimicry: Structural Basis of the CRISPR and Anti-CRISPR Arms Race. Molecular cell 2019, 73, 611-620.e613, doi:10.1016/j.molcel.2018.11.011.

60. Johnston, R.K.; Seamon, K.J.; Saada, E.A.; Podlevsky, J.D.; Branda, S.S.; Timlin, J.A.; Harper, J.C. Use of anti-CRISPR protein AcrIIA4 as a capture ligand for CRISPR/Cas9 detection. Biosensors & bioelectronics 2019, 141, 111361, doi:10.1016/j.bios.2019.111361.

61. Hynes, A.P.; Rousseau, G.M.; Agudelo, D.; Goulet, A.; Amigues, B.; Loehr, J.; Romero, D.A.; Fremaux, C.; Horvath, P.; Doyon, Y.; et al. Widespread anti-CRISPR proteins in virulent bacteriophages inhibit a range of Cas9 proteins. Nature communications 2018, 9, 2919, doi:10.1038/s41467-018-05092-w.

62. Zhu, Y.; Gao, A.; Zhan, Q.; Wang, Y.; Feng, H.; Liu, S.; Gao, G.; Serganov, A.; Gao, P. Diverse Mechanisms of CRISPR-Cas9 Inhibition by Type IIC Anti-CRISPR Proteins. Molecular cell 2019, 74, 296-309.e297, doi:10.1016/j.molcel.2019.01.038.

63. Kim, Y.; Lee, S.J.; Yoon, H.J.; Kim, N.K.; Lee, B.J.; Suh, J.Y. Anti-CRISPR AcrIIC3 discriminates between Cas9 orthologs via targeting the variable surface of the HNH nuclease domain. The FEBS journal 2019, 286, 4661-4674, doi:10.1111/febs.15037.

64. Bubeck, F.; Hoffmann, M.D.; Harteveld, Z.; Aschenbrenner, S.; Bietz, A.; Waldhauer, M.C.; B?rner, K.; Fakhiri, J.; Schmelas, C.; Dietz, L.; et al. Engineered anti-CRISPR proteins for optogenetic control of CRISPR-Cas9. Nature methods 2018, 15, 924-927, doi:10.1038/s41592-018-0178-9.

65. Lee, J.; Mir, A.; Edraki, A.; Garcia, B.; Amrani, N.; Lou, H.E.; Gainetdinov, I.; Pawluk, A.; Ibraheim, R.; Gao, X.D.; et al. Potent Cas9 Inhibition in Bacterial and Human Cells by AcrIIC4 and AcrIIC5 Anti-CRISPR Proteins. mBio 2018, 9, doi:10.1128/mBio.02321-18.

66. Dong, L.; Guan, X.; Li, N.; Zhang, F.; Zhu, Y.; Ren, K.; Yu, L.; Zhou, F.; Han, Z.; Gao, N.; et al. An anti-CRISPR protein disables type V Cas12a by acetylation. Nature structural & molecular biology 2019, 26, 308-314, doi:10.1038/s41594-019-0206-1.

67. Zhang, H.; Li, Z.; Daczkowski, C.M.; Gabel, C.; Mesecar, A.D.; Chang, L. Structural Basis for the Inhibition of CRISPR-Cas12a by Anti-CRISPR Proteins. Cell host & microbe 2019, 25, 815-826.e814, doi:10.1016/j.chom.2019.05.004.

68. Loureiro, A.; da Silva, G.J. CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool. Antibiotics (Basel, Switzerland) 2019, 8, doi:10.3390/antibiotics8010018.

69. Mohanraju, P.; Makarova, K.S.; Zetsche, B.; Zhang, F.; Koonin, E.V.; van der Oost, J. Diverse evolutionary roots and mechanistic variations of the CRISPR-Cas systems. Science (New York, N.Y.) 2016, 353, aad5147, doi:10.1126/science.aad5147.

70. 刁韵; 柴春霞; 王炜. CRISPR/Cas系统在病毒检测中的应用研究进展. 病毒学报. 2023, 39, 1693-1702, doi:10.13242/j.cnki.bingduxuebao.004404.

71. Cameron, P.; Coons, M.M.; Klompe, S.E.; Lied, A.M.; Smith, S.C.; Vidal, B.; Donohoue, P.D.; Rotstein, T.; Kohrs, B.W.; Nyer, D.B.; et al. Harnessing type I CRISPR-Cas systems for genome engineering in human cells. Nature biotechnology 2019, 37, 1471-1477, doi:10.1038/s41587-019-0310-0.

72. Li, J.; Zhao, D.; Zhang, T.; Xiong, H.; Hu, M.; Liu, H.; Zhao, F.; Sun, X.; Fan, P.; Qian, Y.; et al. Precise large-fragment deletions in mammalian cells and mice generated by dCas9-controlled CRISPR/Cas3. Science advances 2024, 10, eadk8052, doi:10.1126/sciadv.adk8052.

73. Cs?rg?, B.; León, L.M.; Chau-Ly, I.J.; Vasquez-Rifo, A.; Berry, J.D.; Mahendra, C.; Crawford, E.D.; Lewis, J.D.; Bondy-Denomy, J. A compact Cascade-Cas3 system for targeted genome engineering. Nature methods 2020, 17, 1183-1190, doi:10.1038/s41592-020-00980-w.

74. Sengupta, A.; Bandyopadhyay, A.; Sarkar, D.; Hendry, J.I.; Schubert, M.G.; Liu, D.; Church, G.M.; Maranas, C.D.; Pakrasi, H.B. Genome streamlining to improve performance of a fast-growing cyanobacterium Synechococcus elongatus UTEX 2973. mBio 2024, 15, e0353023, doi:10.1128/mbio.03530-23.

75. Gomaa, A.A.; Klumpe, H.E.; Luo, M.L.; Selle, K.; Barrangou, R.; Beisel, C.L. Programmable removal of bacterial strains by use of genome-targeting CRISPR-Cas systems. mBio 2014, 5, e00928-00913, doi:10.1128/mBio.00928-13.

76. Selle, K.; Fletcher, J.R.; Tuson, H.; Schmitt, D.S.; McMillan, L.; Vridhambal, G.S.; Rivera, A.J.; Montgomery, S.A.; Fortier, L.C.; Barrangou, R.; et al. In Vivo Targeting of Clostridioides difficile Using Phage-Delivered CRISPR-Cas3 Antimicrobials. mBio 2020, 11, doi:10.1128/mBio.00019-20.

77. Jin, M.; Chen, J.; Zhao, X.; Hu, G.; Wang, H.; Liu, Z.; Chen, W.H. An Engineered λ Phage Enables Enhanced and Strain-Specific Killing of Enterohemorrhagic Escherichia coli. Microbiology spectrum 2022, 10, e0127122, doi:10.1128/spectrum.01271-22.

78. Zhou, Y.; Tang, Y.; Fu, P.; Tian, D.; Yu, L.; Huang, Y.; Li, G.; Li, M.; Wang, Y.; Yang, Z.; et al. The type I-E CRISPR-Cas system influences the acquisition of bla(KPC)-IncF plasmid in Klebsiella pneumonia. Emerging microbes & infections 2020, 9, 1011-1022, doi:10.1080/22221751.2020.1763209.

79. Zhou, Y.; Yang, Y.; Li, X.; Tian, D.; Ai, W.; Wang, W.; Wang, B.; Kreiswirth, B.N.; Yu, F.; Chen, L.; et al. Exploiting a conjugative endogenous CRISPR-Cas3 system to tackle multidrug-resistant Klebsiella pneumoniae. EBioMedicine 2023, 88, 104445, doi:10.1016/j.ebiom.2023.104445.

80. Anzalone, A.V.; Randolph, P.B.; Davis, J.R.; Sousa, A.A.; Koblan, L.W.; Levy, J.M.; Chen, P.J.; Wilson, C.; Newby, G.A.; Raguram, A.; et al. Search-and-replace genome editing without double-strand breaks or donor DNA. Nature 2019, 576, 149-157, doi:10.1038/s41586-019-1711-4.

81. Sun, C.; Lei, Y.; Li, B.; Gao, Q.; Li, Y.; Cao, W.; Yang, C.; Li, H.; Wang, Z.; Li, Y.; et al. Precise integration of large DNA sequences in plant genomes using PrimeRoot editors. Nature biotechnology 2024, 42, 316-327, doi:10.1038/s41587-023-01769-w.

82. Yang, X.; Wilkinson, L.G.; Aubert, M.K.; Houston, K.; Shirley, N.J.; Tucker, M.R. Ovule cell wall composition is a maternal determinant of grain size in barley. The New phytologist 2023, 237, 2136-2147, doi:10.1111/nph.18714.

83. Sha, G.; Sun, P.; Kong, X.; Han, X.; Sun, Q.; Fouillen, L.; Zhao, J.; Li, Y.; Yang, L.; Wang, Y.; et al. Genome editing of a rice CDP-DAG synthase confers multipathogen resistance. Nature 2023, 618, 1017-1023, doi:10.1038/s41586-023-06205-2.

84. Hong, W.J.; Kim, Y.J.; Kim, E.J.; Kumar Nalini Chandran, A.; Moon, S.; Gho, Y.S.; Yoou, M.H.; Kim, S.T.; Jung, K.H. CAFRI-Rice: CRISPR applicable functional redundancy inspector to accelerate functional genomics in rice. The Plant journal : for cell and molecular biology 2020, 104, 532-545, doi:10.1111/tpj.14926.

85. Abdulla, M.F.; Mostafa, K.; Kavas, M. CRISPR/Cas9-mediated mutagenesis of FT/TFL1 in petunia improves plant architecture and early flowering. Plant molecular biology 2024, 114, 69, doi:10.1007/s11103-024-01454-9.

86. Ahmar, S.; Usman, B.; Hensel, G.; Jung, K.H.; Gruszka, D. CRISPR enables sustainable cereal production for a greener future. Trends in plant science 2024, 29, 179-195, doi:10.1016/j.tplants.2023.10.016.

87. Bártová, E. Epigenetic and gene therapy in human and veterinary medicine. Environmental epigenetics 2024, 10, dvae006, doi:10.1093/eep/dvae006.

88. Saunderson, E.A.; Encabo, H.H.; Devis, J.; Rouault-Pierre, K.; Piganeau, M.; Bell, C.G.; Gribben, J.G.; Bonnet, D.; Ficz, G. CRISPR/dCas9 DNA methylation editing is heritable during human hematopoiesis and shapes immune progeny. Proceedings of the National Academy of Sciences of the United States of America 2023, 120, e2300224120, doi:10.1073/pnas.2300224120.

89. Allemailem, K.S.; Alsahli, M.A.; Almatroudi, A.; Alrumaihi, F.; Alkhaleefah, F.K.; Rahmani, A.H.; Khan, A.A. Current updates of CRISPR/Cas9-mediated genome editing and targeting within tumor cells: an innovative strategy of cancer management. Cancer communications (London, England) 2022, 42, 1257-1287, doi:10.1002/cac2.12366.

90. Li, Y.; Li, X.; Qu, J.; Luo, D.; Hu, Z. Cas9 Mediated Correction of β-catenin Mutation and Restoring the Expression of Protein Phosphorylation in Colon Cancer HCT-116 Cells Decrease Cell Proliferation in vitro and Hamper Tumor Growth in Mice in vivo. OncoTargets and therapy 2020, 13, 17-29, doi:10.2147/ott.S225556.

91. Evtimov, V.; Hammett, M.; Nhu-Y, N.; Zhuang, J.; Nisbet, I.; Trounson, A.; Boyd, R.; Shu, R. 12P CRISPR/Cas9-induced knock-out of DGKαζ in TAG-72 CAR-T cells improves function and persistence in ovarian cancer. ESMO Open 2023, 8, doi:10.1016/j.esmoop.2023.100978.

92. Deng, X.; Gao, F.; Li, N.; Li, Q.; Zhou, Y.; Yang, T.; Cai, Z.; Du, P.; Chen, F.; Cai, J. Antitumor activity of NKG2D CAR-T cells against human colorectal cancer cells in vitro and in vivo. American journal of cancer research 2019, 9, 945-958.

93. Gao, L.; Yang, L.; Zhang, S.; Ge, Z.; Su, M.; Shi, Y.; Wang, X.; Huang, C. Engineering NK-92 Cell by Upregulating CXCR2 and IL-2 Via CRISPR-Cas9 Improves Its Antitumor Effects as Cellular Immunotherapy for Human Colon Cancer. Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research 2021, 41, 450-460, doi:10.1089/jir.2021.0078.

94. Zhao, Z.; Shi, L.; Zhang, W.; Han, J.; Zhang, S.; Fu, Z.; Cai, J. CRISPR knock out of programmed cell death protein 1 enhances anti-tumor activity of cytotoxic T lymphocytes. Oncotarget 2018, 9, 5208-5215, doi:10.18632/oncotarget.23730.

95. Zakiyyah, S.N.; Ibrahim, A.U.; Babiker, M.S.; Gaffar, S.; Ozsoz, M.; Zein, M.; Hartati, Y.W. Detection of Tropical Diseases Caused by Mosquitoes Using CRISPR-Based Biosensors. Tropical medicine and infectious disease 2022, 7, doi:10.3390/tropicalmed7100309.

96. Verma, M.K.; Roychowdhury, S.; Sahu, B.D.; Mishra, A.; Sethi, K.K. CRISPR-based point-of-care diagnostics incorporating Cas9, Cas12, and Cas13 enzymes advanced for SARS-CoV-2 detection. Journal of biochemical and molecular toxicology 2022, 36, e23113, doi:10.1002/jbt.23113.

97. Lou, J.; Wang, B.; Li, J.; Ni, P.; Jin, Y.; Chen, S.; Xi, Y.; Zhang, R.; Duan, G. The CRISPR-Cas system as a tool for diagnosing and treating infectious diseases. Molecular biology reports 2022, 49, 11301-11311, doi:10.1007/s11033-022-07752-z.

98. Kostyusheva, A.; Brezgin, S.; Babin, Y.; Vasilyeva, I.; Glebe, D.; Kostyushev, D.; Chulanov, V. CRISPR-Cas systems for diagnosing infectious diseases. Methods (San Diego, Calif.) 2022, 203, 431-446, doi:10.1016/j.ymeth.2021.04.007.

99. Shinoda, H.; Taguchi, Y.; Nakagawa, R.; Makino, A.; Okazaki, S.; Nakano, M.; Muramoto, Y.; Takahashi, C.; Takahashi, I.; Ando, J.; et al. Amplification-free RNA detection with CRISPR-Cas13. Communications biology 2021, 4, 476, doi:10.1038/s42003-021-02001-8.

100. Liu, P.; Wang, X.; Liang, J.; Dong, Q.; Zhang, J.; Liu, D.; Wang, S.; Bi, J.; Liu, W.; Wang, Z.; et al. A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosis. Frontiers in microbiology 2021, 12, 796916, doi:10.3389/fmicb.2021.796916.

101. Chen, Y.; Mei, Y.; Zhao, X.; Jiang, X. Reagents-Loaded, Automated Assay that Integrates Recombinase-Aided Amplification and Cas12a Nucleic Acid Detection for a Point-of-Care Test. Analytical chemistry 2020, 92, 14846-14852, doi:10.1021/acs.analchem.0c03883.

102. Leta, S.; Chibssa, T.R.; Paeshuyse, J. CRISPR-Cas12/Cas13: Bibliometric analysis and systematic review of its application in infectious disease detection. Journal of infection and public health 2024, 17, 741-747, doi:10.1016/j.jiph.2024.03.003.

103. Dai, Y.; Somoza, R.A.; Wang, L.; Welter, J.F.; Li, Y.; Caplan, A.I.; Liu, C.C. Exploring the Trans-Cleavage Activity of CRISPR-Cas12a (cpf1) for the Development of a Universal Electrochemical Biosensor. Angewandte Chemie (International ed. in English) 2019, 58, 17399-17405, doi:10.1002/anie.201910772.

104. Yue, Y.; Wang, S.; Jin, Q.; An, N.; Wu, L.; Huang, H. A triple amplification strategy using GR-5 DNAzyme as a signal medium for ultrasensitive detection of trace Pb(2+) based on CRISPR/Cas12a empowered electrochemical biosensor. Analytica chimica acta 2023, 1263, 341241, doi:10.1016/j.aca.2023.341241.

105. Vuono, E.A.; Ramirez-Medina, E.; Pruitt, S.; Rai, A.; Espinoza, N.; Velazquez-Salinas, L.; Gladue, D.P.; Borca, M.V. Evaluation of the Function of the ASFV KP177R Gene, Encoding for Structural Protein p22, in the Process of Virus Replication and in Swine Virulence. Viruses 2021, 13, doi:10.3390/v13060986.

106. Luan, H.; Wang, S.; Ju, L.; Liu, T.; Shi, H.; Ge, S.; Jiang, S.; Wu, J.; Peng, J. KP177R-based visual assay integrating RPA and CRISPR/Cas12a for the detection of African swine fever virus. Frontiers in immunology 2024, 15, 1358960, doi:10.3389/fimmu.2024.1358960.

107. Bu, S.; Liu, X.; Wang, Z.; Wei, H.; Yu, S.; Li, Z.; Hao, Z.; Liu, W.; Wan, J. Ultrasensitive detection of pathogenic bacteria by CRISPR/Cas12a coupling with a primer exchange reaction. Sensors and Actuators B: Chemical 2021, 347, doi:10.1016/j.snb.2021.130630.

108. Wu, C.; Wang, X.; Guo, L.; Huang, X.; Wu, L.; Huang, H. An electrochemical aptasensor based on exonuclease III-assisted signal amplification coupled with CRISPR-Cas12a for ochratoxin A detection. Food Control 2023, 148, doi:10.1016/j.foodcont.2023.109631.

109. Huang, L.; Yuan, N.; Guo, W.; Zhang, Y.; Zhang, W. An electrochemical biosensor for the highly sensitive detection of Staphylococcus aureus based on SRCA-CRISPR/Cas12a. Talanta 2023, 252, 123821, doi:10.1016/j.talanta.2022.123821.

110. Burstein, D.; Harrington, L.B.; Strutt, S.C.; Probst, A.J.; Anantharaman, K.; Thomas, B.C.; Doudna, J.A.; Banfield, J.F. New CRISPR-Cas systems from uncultivated microbes. Nature 2017, 542, 237-241, doi:10.1038/nature21059.

111. 贺金荣; 张渝疆; 王宇萌; 蔡虹; 张志凯; 李伟; 梁莹. 新疆地区58株鼠疫耶尔森菌规律聚集的间隔短回文重复位点多态性分析. 中国媒介生物学及控制杂志. 2017, 28, 233-237.

112. Zhang, M.M.; Wong, F.T.; Wang, Y.; Luo, S.; Lim, Y.H.; Heng, E.; Yeo, W.L.; Cobb, R.E.; Enghiad, B.; Ang, E.L.; et al. CRISPR-Cas9 strategy for activation of silent Streptomyces biosynthetic gene clusters. Nature chemical biology 2017, doi:10.1038/nchembio.2341.

113. Ahmad, A.; Jamil, A.; Munawar, N. GMOs or non-GMOs? The CRISPR Conundrum. Frontiers in plant science 2023, 14, 1232938, doi:10.3389/fpls.2023.1232938.

基本信息:

中图分类号:Q78

引用信息:

[1]卢泽宇,李蕴华,李媛,等.CRISPR-Cas系统的多样化类型、机制及其应用进展[J].经济动物学报().

基金信息:

内蒙古自治区科技计划项目(2023YFHH0076); 国家自然科学基金项目(32160772); 内蒙古自治区自然科学基金项目(2024ZD14); 内蒙古自治区高等学校创新团队发展计划项目(NMGIRT2322)

发布时间:

2025-04-16

出版时间:

2025-04-16

网络发布时间:

2025-04-16

检 索 高级检索