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A New Class of Medicines through DNA Editing

Matthew H. Porteus ... 肿瘤 • 2019.03.07
• CRISPR-Cas9基因编辑技术治疗镰状细胞病和β地中海贫血 • 通过BCL11A转录后基因沉默策略治疗镰状细胞病 • 风暴过后——探索负责任的基因组编辑之路 • 修复人类生殖细胞系的有力理由



Matthew H. Porteus, M.D., Ph.D.
From the Department of Pediatrics–Stem Cell Transplantation, Stanford University, Stanford, CA. Address reprint requests to Dr. Porteus at the Department of Pediatrics, Lorry Lokey Stem Cell Research Bldg. MC 5462, 265 Campus Dr., Stanford CA 94305, or at mporteus@stanford.edu.



1. 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;317:230-234.

2. Rouet P, Smih F, Jasin M. Introduction of double-strand breaks into the genome of mouse cells by expression of a rare-cutting endonuclease. Mol Cell Biol 1994;14:8096-8106.

3. Rouet P, Smih F, Jasin M. Expression of a site-specific endonuclease stimulates homologous recombination in mammalian cells. Proc Natl Acad Sci U S A 1994;91:6064-6068.

4. Porteus MH, Baltimore D. Chimeric nucleases stimulate gene targeting in human cells. Science 2003;300:763-763.

5. Choulika A, Perrin A, Dujon B, Nicolas J-F. Induction of homologous recombination in mammalian chromosomes by using the I-SceI system of Saccharomyces cerevisiae. Mol Cell Biol 1995;15:1968-1973.

6. Bibikova M, Golic M, Golic KG, Carroll D. Targeted chromosomal cleavage and mutagenesis in Drosophila using zinc-finger nucleases. Genetics 2002;161:1169-1175.

7. Carroll D. Genome engineering with targetable nucleases. Annu Rev Biochem 2014;83:409-439.

8. Porteus MH, Carroll D. Gene targeting using zinc finger nucleases. Nat Biotechnol 2005;23:967-973.

9. Urnov FD, Miller JC, Lee YL, et al. Highly efficient endogenous human gene correction using designed zinc-finger nucleases. Nature 2005;435:646-651.

10. Bibikova M, Beumer K, Trautman JK, Carroll D. Enhancing gene targeting with designed zinc finger nucleases. Science 2003;300:764-764.

11. Durai S, Mani M, Kandavelou K, Wu J, Porteus MH, Chandrasegaran S. Zinc finger nucleases: custom-designed molecular scissors for genome engineering of plant and mammalian cells. Nucleic Acids Res 2005;33:5978-5990.

12. Porteus M. Genome editing: a new approach to human therapeutics. Annu Rev Pharmacol Toxicol 2016;56:163-190.

13. Russell DW, Hirata RK. Human gene targeting by viral vectors. Nat Genet 1998;18:325-330.

14. Ricciardi AS, Quijano E, Putman R, Saltzman WM, Glazer PM. Peptide nucleic acids as a tool for site-specific gene editing. Molecules 2018;23(3):E632-E632.

15. Komor AC, Kim YB, Packer MS, Zuris JA, Liu DR. Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature 2016;533:420-424.

16. Gaudelli NM, Komor AC, Rees HA, et al. Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 2017;551:464-471.

17. Barrangou R, Horvath P. A decade of discovery: CRISPR functions and applications. Nat Microbiol 2017;2:17092-17092.

18. Horvath P, Barrangou R. CRISPR/Cas, the immune system of bacteria and archaea. Science 2010;327:167-170.

19. Doudna JA, Charpentier E. Genome editing: the new frontier of genome engineering with CRISPR-Cas9. Science 2014;346:1258096-1258096.

20. Hsu PD, Lander ES, Zhang F. Development and applications of CRISPR-Cas9 for genome engineering. Cell 2014;157:1262-1278.

21. West SC, Chappell C, Hanakahi LA, Masson JY, McIlwraith MJ, Van Dyck E. Double-strand break repair in human cells. Cold Spring Harb Symp Quant Biol 2000;65:315-321.

22. Kanaar R, Hoeijmakers JH, van Gent DC. Molecular mechanisms of DNA double strand break repair. Trends Cell Biol 1998;8:483-489.

23. Hendel A, Bak RO, Clark JT, et al. Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. Nat Biotechnol 2015;33:985-989.

24. Bétermier M, Bertrand P, Lopez BS. Is non-homologous end-joining really an inherently error-prone process? PLoS Genet 2014;10(1):e1004086-e1004086.

25. Lin Y, Waldman AS. Capture of DNA sequences at double-strand breaks in mammalian chromosomes. Genetics 2001;158:1665-1674.

26. Hendel A, Kildebeck EJ, Fine EJ, et al. Quantifying genome-editing outcomes at endogenous loci with SMRT sequencing. Cell Rep 2014;7:293-305.

27. Li H, Haurigot V, Doyon Y, et al. In vivo genome editing restores haemostasis in a mouse model of haemophilia. Nature 2011;475:217-221.

28. Nakade S, Tsubota T, Sakane Y, et al. Microhomology-mediated end-joining-dependent integration of donor DNA in cells and animals using TALENs and CRISPR/Cas9. Nat Commun 2014;5:5560-5560.

29. Suzuki K, Tsunekawa Y, Hernandez-Benitez R, et al. In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration. Nature 2016;540:144-149.

30. Qasim W, Zhan H, Samarasinghe S, et al. Molecular remission of infant B-ALL after infusion of universal TALEN gene-edited CAR T cells. Sci Transl Med 2017;9(374):eaaj2013-eaaj2013.

31. Bak RO, Dever DP, Reinisch A, Cruz Hernandez D, Majeti R, Porteus MH. Multiplexed genetic engineering of human hematopoietic stem and progenitor cells using CRISPR/Cas9 and AAV6. Elife 2017;6:e27873-e27873.

32. Chen F, Pruett-Miller SM, Huang Y, et al. High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases. Nat Methods 2011;8:753-755.

33. Richardson CD, Kazane KR, Feng SJ, et al. CRISPR-Cas9 genome editing in human cells works via the Fanconi Anemia pathway. bioRxiv. May 9, 2017 (https://www.biorxiv.org/content/early/2017/05/09/136028).

34. Dever DP, Bak RO, Reinisch A, et al. CRISPR/Cas9 β-globin gene targeting in human haematopoietic stem cells. Nature 2016;539:384-389.

35. DeWitt MA, Magis W, Bray NL, et al. Selection-free genome editing of the sickle mutation in human adult hematopoietic stem/progenitor cells. Sci Transl Med 2016;8:360ra134-360ra134.

36. De Ravin SS, Li L, Wu X, et al. CRISPR-Cas9 gene repair of hematopoietic stem cells from patients with X-linked chronic granulomatous disease. Sci Transl Med 2017;9(372):eaah3480-eaah3480.

37. Schiroli G, Ferrari S, Conway A, et al. Preclinical modeling highlights the therapeutic potential of hematopoietic stem cell gene editing for correction of SCID-X1. Sci Transl Med 2017;9(411):eaan0820-eaan0820.

38. Hubbard N, Hagin D, Sommer K, et al. Targeted gene editing restores regulated CD40L function in X-linked hyper-IgM syndrome. Blood 2016;127:2513-2522.

39. Eyquem J, Mansilla-Soto J, Giavridis T, et al. Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection. Nature 2017;543:113-117.

40. Lombardo A, Cesana D, Genovese P, et al. Site-specific integration and tailoring of cassette design for sustainable gene transfer. Nat Methods 2011;8:861-869.

41. DiGiusto DL, Cannon PM, Holmes MC, et al. Preclinical development and qualification of ZFN-mediated CCR5 disruption in human hematopoietic stem/progenitor cells. Mol Ther Methods Clin Dev 2016;3:16067-16067.

42. Cromer MK, Vaidyanathan S, Ryan DE, et al. Global transcriptional response to CRISPR/Cas9-AAV6-based genome editing in CD34+ hematopoietic stem and progenitor cells. Mol Ther 2018;26:2431-2442.

43. Ihry RJ, Worringer KA, Salick MR, et al. p53 Inhibits CRISPR-Cas9 engineering in human pluripotent stem cells. Nat Med 2018;24:939-946.

44. Wang J, Exline CM, DeClercq JJ, et al. Homology-driven genome editing in hematopoietic stem and progenitor cells using ZFN mRNA and AAV6 donors. Nat Biotechnol 2015;33:1256-1263.

45. Sather BD, Romano Ibarra GS, Sommer K, et al. Efficient modification of CCR5 in primary human hematopoietic cells using a megaTAL nuclease and AAV donor template. Sci Transl Med 2015;7:307ra156-307ra156.

46. De Ravin SS, Reik A, Liu PQ, et al. Targeted gene addition in human CD34(+) hematopoietic cells for correction of X-linked chronic granulomatous disease. Nat Biotechnol 2016;34:424-429.

47. Bak RO, Dever DP, Porteus MH. CRISPR/Cas9 genome editing in human hematopoietic stem cells. Nat Protoc 2018;13:358-376.

48. Certo MT, Ryu BY, Annis JE, et al. Tracking genome engineering outcome at individual DNA breakpoints. Nat Methods 2011;8:671-676.

49. Potts PR, Porteus MH, Yu H. Human SMC5/6 complex promotes sister chromatid homologous recombination by recruiting the SMC1/3 cohesin complex to double-strand breaks. EMBO J 2006;25:3377-3388.

50. Canny MD, Moatti N, Wan LCK, et al. Inhibition of 53BP1 favors homology-dependent DNA repair and increases CRISPR-Cas9 genome-editing efficiency. Nat Biotechnol 2018;36:95-102.

51. Paulsen BS, Mandal PK, Frock RL, et al. Ectopic expression of RAD52 and dn53BP1 improves homology-directed repair during CRISPR–Cas9 genome editing. Nat Biomed Eng 2017;1:878-888 (https://www.nature.com/articles/s41551-017-0145-2).

52. Sharma R, Anguela XM, Doyon Y, et al. In vivo genome editing of the albumin locus as a platform for protein replacement therapy. Blood 2015;126:1777-1784.

53. Holt N, Wang J, Kim K, et al. Human hematopoietic stem/progenitor cells modified by zinc-finger nucleases targeted to CCR5 control HIV-1 in vivo. Nature Biotechnol 2010;28:839-847.

54. Perez EE, Wang J, Miller JC, et al. Establishment of HIV-1 resistance in CD4+ T cells by genome editing using zinc-finger nucleases. Nat Biotechnol 2008;26:808-816.

55. Tebas P, Stein D, Tang WW, et al. Gene editing of CCR5 in autologous CD4 T cells of persons infected with HIV. N Engl J Med 2014;370:901-910.

56. Poirot L, Philip B, Schiffer-Mannioui C, et al. Multiplex genome-edited T-cell manufacturing platform for “off-the-shelf” adoptive T-cell immunotherapies. Cancer Res 2015;75:3853-3864.

57. Hu Z, Ding W, Zhu D, et al. TALEN-mediated targeting of HPV oncogenes ameliorates HPV-related cervical malignancy. J Clin Invest 2015;125:425-436.

58. Cyranoski D. CRISPR gene-editing tested in a person for the first time. Nature 2016;539:479-479.

59. Canver MC, Smith EC, Sher F, et al. BCL11A enhancer dissection by Cas9-mediated in situ saturating mutagenesis. Nature 2015;527:192-197.

60. Voit RA, McMahon MA, Sawyer SL, Porteus MH. Generation of an HIV resistant T-cell line by targeted “stacking” of restriction factors. Mol Ther 2013;21:786-795.

61. Wang L, Smith J, Breton C, et al. Meganuclease targeting of PCSK9 in macaque liver leads to stable reduction in serum cholesterol. Nat Biotechnol 2018;36:717-725.

62. Tabebordbar M, Zhu K, Cheng JKW, et al. In vivo gene editing in dystrophic mouse muscle and muscle stem cells. Science 2016;351:407-411.

63. Long C, Amoasii L, Mireault AA, et al. Postnatal genome editing partially restores dystrophin expression in a mouse model of muscular dystrophy. Science 2016;351:400-403.

64. Nelson CE, Hakim CH, Ousterout DG, et al. In vivo genome editing improves muscle function in a mouse model of Duchenne muscular dystrophy. Science 2016;351:403-407.

65. Yin H, Xue W, Chen S, et al. Genome editing with Cas9 in adult mice corrects a disease mutation and phenotype. Nat Biotechnol 2014;32:551-553.

66. Yang Y, Wang L, Bell P, et al. A dual AAV system enables the Cas9-mediated correction of a metabolic liver disease in newborn mice. Nat Biotechnol 2016;34:334-338.

67. Charlesworth CT, Deshpande PS, Dever DP, et al. Identification of pre-existing adaptive immunity to Cas9 proteins in humans. bioRxiv January 5, 2018 (https://www.biorxiv.org/content/early/2018/01/05/243345).

68. Wagner DL, Amini L, Wendering DJ, Reinke P, Volk H-P, Schmueck-Henneresse M. High prevalence of S. pyogenes Cas9-specific T cell sensitization within the adult human population — a balanced effector/regulatory T cell response. bioRxiv. April 4, 2018 (https://www.biorxiv.org/content/early/2018/04/04/295139).

69. Tsai SQ, Joung JK. Defining and improving the genome-wide specificities of CRISPR-Cas9 nucleases. Nat Rev Genet 2016;17:300-312.

70. Chen JS, Dagdas YS, Kleinstiver BP, et al. Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature 2017;550:407-410.

71. Casini A, Olivieri M, Petris G, et al. A highly specific SpCas9 variant is identified by in vivo screening in yeast. Nat Biotechnol 2018;36:265-271.

72. Vakulskas CA, Dever DP, Rettig GR, et al. A high-fidelity Cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human hematopoietic stem and progenitor cells. Nat Med 2018;24:1216-1224.

73. Benabdallah BF, Allard E, Yao S, et al. Targeted gene addition to human mesenchymal stromal cells as a cell-based plasma-soluble protein delivery platform. Cytotherapy 2010;12:394-399.

74. Barker JC, Barker AD, Bills J, et al. Genome editing of mouse fibroblasts by homologous recombination for sustained secretion of PDGF-B and augmentation of wound healing. Plast Reconstr Surg 2014;134(3):389e-401e.

75. National Academies of Sciences, Engineering, and Medicine. Human genome editing: science, ethics, and governance. Washington, DC: National Academies Press, 2017.

76. Genome editing and human reproduction: social and ethical issues. London: Nuffield Council on Bioethics, 2018 (http://nuffieldbioethics.org/project/genome-editing-human-reproduction). 

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