Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Review Article
  • Published:

Genome editing using CRISPR/Cas9 to treat hereditary hematological disorders

Gene Therapy volume 29, pages 207–216 (2022)Cite this article

Subjects

Abstract

The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system is a versatile and convenient genome-editing tool with prospects in gene therapy. This technique is based on customized site-specific nucleases with programmable guiding RNAs that cleave and introduce double-strand breaks (DSBs) at the target locus and achieve precise genome modification by triggering DNA repair mechanisms. Human hematopoietic stem/progenitor cells (HSPCs) are conventional cell targets for gene therapy in hematological diseases and have been widely used in most studies. Induced pluripotent stem cells (iPSCs) can be generated from a variety of somatic cells and hold great promise for personalized cell-based therapies. CRISPR/Cas9-mediated genome editing in autologous HSPCs and iPSCs is an ideal therapeutic solution for treating hereditary hematological disorders. Here, we review and summarize the latest studies about CRISPR/Cas9-mediated genome editing in patient-derived HSPCs and iPSCs to treat hereditary hematological disorders. Current challenges and prospects are also discussed.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Overview of CRISPR/Cas9-mediated therapeutic genome editing in hereditary hematological disorders.

Similar content being viewed by others

References

  1. 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.

    Article  CAS  PubMed  Google Scholar 

  2. 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.

    Article  CAS  PubMed  Google Scholar 

  3. 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.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F. Genome engineering using the CRISPR-Cas9 system. Nat Protocols. 2013;8:2281–308.

    Article  CAS  PubMed  Google Scholar 

  5. Mao Z, Bozzella M, Seluanov A, Gorbunova V. DNA repair by nonhomologous end joining and homologous recombination during cell cycle in human cells. Cell Cycle (Georgetown. Tex). 2008;7:2902–6.

    Article  CAS  Google Scholar 

  6. Ribeil JA, Hacein-Bey-Abina S, Payen E, Magnani A, Semeraro M, Magrin E, et al. Gene therapy in a patient with sickle cell disease. N Engl J Med. 2017;376:848–55.

    Article  CAS  PubMed  Google Scholar 

  7. Thompson AA, Walters MC, Kwiatkowski J, Rasko JEJ, Ribeil JA, Hongeng S, et al. Gene therapy in patients with transfusion-dependent β-thalassemia. N Engl J Med. 2018;378:1479–93.

    Article  CAS  PubMed  Google Scholar 

  8. Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006;126:663–76.

    Article  CAS  PubMed  Google Scholar 

  9. Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007;131:861–72.

    Article  CAS  PubMed  Google Scholar 

  10. Papapetrou EP. Induced pluripotent stem cells, past and future. Science. 2016;353:991–2.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Hanna J, Wernig M, Markoulaki S, Sun CW, Meissner A, Cassady JP, et al. Treatment of sickle cell anemia mouse model with iPS cells generated from autologous skin. Science. 2007;318:1920–3.

    Article  CAS  PubMed  Google Scholar 

  12. Zhou H, Wu S, Joo JY, Zhu S, Han DW, Lin T, et al. Generation of induced pluripotent stem cells using recombinant proteins. Cell Stem Cell. 2009;4:381–4.

    Article  CAS  PubMed  Google Scholar 

  13. Jia F, Wilson KD, Sun N, Gupta DM, Huang M, Li Z, et al. A nonviral minicircle vector for deriving human iPS cells. Nat Methods. 2010;7:197–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Warren L, Manos PD, Ahfeldt T, Loh YH, Li H, Lau F, et al. Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA. Cell Stem Cell. 2010;7:618–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Nishimura K, Sano M, Ohtaka M, Furuta B, Umemura Y, Nakajima Y, et al. Development of defective and persistent Sendai virus vector: a unique gene delivery/expression system ideal for cell reprogramming. J Biol Chem. 2011;286:4760–71.

    Article  CAS  PubMed  Google Scholar 

  16. Martin RM, Ikeda K, Cromer MK, Uchida N, Nishimura T, Romano R, et al. Highly efficient and marker-free genome editing of human pluripotent stem cells by CRISPR-Cas9 RNP and AAV6 donor-mediated homologous recombination. Cell Stem Cell. 2019;24:821–8.e5.

    Article  CAS  PubMed  Google Scholar 

  17. Hu Z, Zhou M, Wu Y, Li Z, Liu X, Wu L, et al. ssODN-mediated in-frame deletion with CRISPR/Cas9 Restores FVIII function in hemophilia a-patient-derived iPSCs and ECs. Mol Ther Nucleic Acids. 2019;17:198–209.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Morishige S, Mizuno S, Ozawa H, Nakamura T, Mazahery A, Nomura K, et al. CRISPR/Cas9-mediated gene correction in hemophilia B patient-derived iPSCs. Int J Hematol. 2020;111:225–33.

    Article  CAS  PubMed  Google Scholar 

  19. Xie F, Ye L, Chang JC, Beyer AI, Wang J, Muench MO, et al. Seamless gene correction of β-thalassemia mutations in patient-specific iPSCs using CRISPR/Cas9 and piggyBac. Genome Res. 2014;24:1526–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Wang Y, Zheng CG, Jiang Y, Zhang J, Chen J, Yao C, et al. Genetic correction of β-thalassemia patient-specific iPS cells and its use in improving hemoglobin production in irradiated SCID mice. Cell Res. 2012;22:637–48.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Zwaka TP, Thomson JA. Homologous recombination in human embryonic stem cells. Nat Biotechnol. 2003;21:319–21.

    Article  CAS  PubMed  Google Scholar 

  22. Ma N, Liao B, Zhang H, Wang L, Shan Y, Xue Y, et al. Transcription activator-like effector nuclease (TALEN)-mediated gene correction in integration-free β-thalassemia induced pluripotent stem cells. J Biol Chem. 2013;288:34671–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Sun N, Zhao H. Seamless correction of the sickle cell disease mutation of the HBB gene in human induced pluripotent stem cells using TALENs. Biotechnol Bioeng. 2014;111:1048–53.

    Article  CAS  PubMed  Google Scholar 

  24. Hoban MD, Cost GJ, Mendel MC, Romero Z, Kaufman ML, Joglekar AV, et al. Correction of the sickle cell disease mutation in human hematopoietic stem/progenitor cells. Blood. 2015;125:2597–604.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, et al. Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Nature. 2018;556:57–63.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Nishimasu H, Shi X, Ishiguro S, Gao L, Hirano S, Okazaki S, et al. Engineered CRISPR-Cas9 nuclease with expanded targeting space. Science. 2018;361:1259–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Chatterjee P, Lee J, Nip L, Koseki SRT, Tysinger E, Sontheimer EJ, et al. A Cas9 with PAM recognition for adenine dinucleotides. Nat Commun. 2020;11:2474.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Walton RT, Christie KA, Whittaker MN, Kleinstiver BP. Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Science. 2020;368:290–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, et al. Multiplex genome engineering using CRISPR/Cas systems. Science. 2013;339:819–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Doudna JA. The promise and challenge of therapeutic genome editing. Nature. 2020;578:229–36.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Dever DP, Bak RO, Reinisch A, Camarena J, Washington G, Nicolas CE, et al. CRISPR/Cas9 β-globin gene targeting in human haematopoietic stem cells. Nature. 2016;539:384–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Taher AT, Weatherall DJ, Cappellini MD. Thalassaemia. Lancet. 2018;391:155–67.

    Article  PubMed  Google Scholar 

  33. Gaziev J, Lucarelli G. Allogeneic cellular gene therapy for hemoglobinopathies. Hematol Oncol Clin North Am. 2010;24:1145–63.

    Article  PubMed  Google Scholar 

  34. Vakulskas CA, Dever DP, Rettig GR, Turk R, Jacobi AM, Collingwood MA, 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;24:1216–24.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Kountouris P, Lederer CW, Fanis P, Feleki X, Old J, Kleanthous M. IthaGenes: an interactive database for haemoglobin variations and epidemiology. PLoS ONE. 2014;9:e103020.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  36. Charlesworth CT, Camarena J, Cromer MK, Vaidyanathan S, Bak RO, Carte JM, et al. Priming human repopulating hematopoietic stem and progenitor cells for Cas9/sgRNA Gene Targeting. Mol Ther Nucleic Acids. 2018;12:89–104.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Yang Y, Zhang X, Yi L, Hou Z, Chen J, Kou X, et al. Naïve induced pluripotent stem cells generated from β-thalassemia fibroblasts allow efficient gene correction with CRISPR/Cas9. Stem Cells Transl Med. 2016;5:8–19.

    Article  CAS  PubMed  Google Scholar 

  38. Liu Y, Yang Y, Kang X, Lin B, Yu Q, Song B, et al. One-step biallelic and scarless correction of a β-Thalassemia mutation in patient-specific iPSCs without drug selection. Mol Ther Nucleic Acids. 2017;6:57–67.

    Article  CAS  PubMed  Google Scholar 

  39. Cai L, Bai H, Mahairaki V, Gao Y, He C, Wen Y, et al. A universal approach to correct various HBB gene mutations in human stem cells for gene therapy of beta-thalassemia and sickle cell disease. Stem Cells Transl Med. 2018;7:87–97.

    Article  CAS  PubMed  Google Scholar 

  40. Ware RE, de Montalembert M, Tshilolo L, Abboud MR. Sickle cell disease. Lancet. 2017;390:311–23.

    Article  PubMed  Google Scholar 

  41. Park SH, Lee CM, Dever DP, Davis TH, Camarena J, Srifa W, et al. Highly efficient editing of the β-globin gene in patient-derived hematopoietic stem and progenitor cells to treat sickle cell disease. Nucleic Acids Res. 2019;47:7955–72.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Haro-Mora JJ, Uchida N, Demirci S, Wang Q, Zou J, Tisdale JF. Biallelic correction of sickle cell disease-derived induced pluripotent stem cells (iPSCs) confirmed at the protein level through serum-free iPS-sac/erythroid differentiation. Stem Cells Transl Med. 2020;9:590–602.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Park S, Gianotti-Sommer A, Molina-Estevez FJ, Vanuytsel K, Skvir N, Leung A, et al. A comprehensive, ethnically diverse library of sickle cell disease-specific induced pluripotent stem cells. Stem Cell Rep. 2017;8:1076–85.

    Article  CAS  Google Scholar 

  44. Martyn GE, Wienert B, Yang L, Shah M, Norton LJ, Burdach J, et al. Natural regulatory mutations elevate the fetal globin gene via disruption of BCL11A or ZBTB7A binding. Nat Genet. 2018;50:498–503.

    Article  CAS  PubMed  Google Scholar 

  45. Wu Y, Zeng J, Roscoe BP, Liu P, Yao Q, Lazzarotto CR, et al. Highly efficient therapeutic gene editing of human hematopoietic stem cells. Nat Med. 2019;25:776–83.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Traxler EA, Yao Y, Wang YD, Woodard KJ, Kurita R, Nakamura Y, et al. A genome-editing strategy to treat β-hemoglobinopathies that recapitulates a mutation associated with a benign genetic condition. Nat Med. 2016;22:987–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Antoniani C, Meneghini V, Lattanzi A, Felix T, Romano O, Magrin E, et al. Induction of fetal hemoglobin synthesis by CRISPR/Cas9-mediated editing of the human β-globin locus. Blood. 2018;131:1960–73.

    Article  CAS  PubMed  Google Scholar 

  48. Frangoul H, Altshuler D, Cappellini MD, Chen YS, Domm J, Eustace BK, et al. CRISPR-Cas9 gene editing for sickle cell disease and β-thalassemia. N Engl J Med. 2021;384:252–60.

    Article  CAS  PubMed  Google Scholar 

  49. Srivastava A, Brewer AK, Mauser-Bunschoten EP, Key NS, Kitchen S, Llinas A, et al. Guidelines for the management of hemophilia. Haemophilia. 2013;19:e1–47.

    Article  CAS  PubMed  Google Scholar 

  50. Nathwani AC, Reiss UM, Tuddenham EG, Rosales C, Chowdary P, McIntosh J, et al. Long-term safety and efficacy of factor IX gene therapy in hemophilia B. N Engl J Med. 2014;371:1994–2004.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  51. Rangarajan S, Walsh L, Lester W, Perry D, Madan B, Laffan M, et al. AAV5-factor VIII gene transfer in severe hemophilia A.N Engl J Med. 2017;377:2519–30.

    Article  CAS  PubMed  Google Scholar 

  52. Pavani G, Laurent M, Fabiano A, Cantelli E, Sakkal A, Corre G, et al. Ex vivo editing of human hematopoietic stem cells for erythroid expression of therapeutic proteins. Nat Commun. 2020;11:3778.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Park CY, Kim DH, Son JS, Sung JJ, Lee J, Bae S, et al. Functional correction of large factor VIII gene chromosomal inversions in hemophilia a patient-derived iPSCs using CRISPR-Cas9. Cell Stem Cell. 2015;17:213–20.

    Article  CAS  PubMed  Google Scholar 

  54. Payne AB, Miller CH, Kelly FM, Michael Soucie J, Craig, Hooper W. The CDC hemophilia A autation project (CHAMP) mutation list: a new online resource. Hum Mutat. 2013;34:E2382–91.

    Article  CAS  PubMed  Google Scholar 

  55. Sung JJ, Park CY, Leem JW, Cho MS, Kim DW. Restoration of FVIII expression by targeted gene insertion in the FVIII locus in hemophilia A patient-derived iPSCs. Exp Mol Med. 2019;51:1–9.

    Article  CAS  PubMed  Google Scholar 

  56. Park CY, Sung JJ, Cho SR, Kim J, Kim DW. Universal correction of blood coagulation factor VIII in patient-derived induced pluripotent stem cells using CRISPR/Cas9. Stem Cell Rep. 2019;12:1242–9.

    Article  CAS  Google Scholar 

  57. Li T, Miller CH, Payne AB, Craig, Hooper W. The CDC hemophilia B mutation project mutation list: a new online resource. Mol Genet Genomic Med. 2013;1:238–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Lyu C, Shen J, Wang R, Gu H, Zhang J, Xue F, et al. Targeted genome engineering in human induced pluripotent stem cells from patients with hemophilia B using the CRISPR-Cas9 system. Stem Cell Res Therapy. 2018;9:92.

    Article  CAS  Google Scholar 

  59. Ramaswamy S, Tonnu N, Menon T, Lewis BM, Green KT, Wampler D, et al. Autologous and heterologous cell therapy for hemophilia B toward functional restoration of factor IX. Cell Rep. 2018;23:1565–80.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Fischer A, Notarangelo LD, Neven B, Cavazzana M, Puck JM. Severe combined immunodeficiencies and related disorders. Nat Rev Dis Primers. 2015;1:15061.

    Article  PubMed  Google Scholar 

  61. Pavel-Dinu M, Wiebking V, Dejene BT, Srifa W, Mantri S, Nicolas CE, et al. Gene correction for SCID-X1 in long-term hematopoietic stem cells. Nat Commun. 2019;10:1634.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  62. Chang CW, Lai YS, Westin E, Khodadadi-Jamayran A, Pawlik KM, Lamb LS Jr., et al. Modeling human severe combined immunodeficiency and correction by CRISPR/Cas9-enhanced gene targeting. Cell Rep. 2015;12:1668–77.

    Article  CAS  PubMed  Google Scholar 

  63. Koyanagi-Aoi M, Ohnuki M, Takahashi K, Okita K, Noma H, Sawamura Y, et al. Differentiation-defective phenotypes revealed by large-scale analyses of human pluripotent stem cells. Proc Natl Acad Sci USA. 2013;110:20569–74.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. Takahashi K, Yamanaka S. A decade of transcription factor-mediated reprogramming to pluripotency. Nat Rev Mol Cell Biol. 2016;17:183–93.

    Article  CAS  PubMed  Google Scholar 

  65. Lin S, Staahl BT, Alla RK, Doudna JA. Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery. eLife. 2014;3:e04766.

    Article  PubMed  PubMed Central  Google Scholar 

  66. Maruyama T, Dougan SK, Truttmann MC, Bilate AM, Ingram JR, Ploegh HL. Increasing the efficiency of precise genome editing with CRISPR-Cas9 by inhibition of nonhomologous end joining. Nat Biotechnol. 2015;33:538–42.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  67. Chu VT, Weber T, Wefers B, Wurst W, Sander S, Rajewsky K, et al. Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nat Biotechnol. 2015;33:543–8.

    Article  CAS  PubMed  Google Scholar 

  68. Hendel A, Bak RO, Clark JT, Kennedy AB, Ryan DE, Roy S, et al. Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. Nat Biotechnol. 2015;33:985–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  69. Richardson CD, Ray GJ, DeWitt MA, Curie GL, Corn JE. Enhancing homology-directed genome editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA. Nat Biotechnol. 2016;34:339–44.

    Article  CAS  PubMed  Google Scholar 

  70. O’Driscoll M, Cerosaletti KM, Girard PM, Dai Y, Stumm M, Kysela B, et al. DNA ligase IV mutations identified in patients exhibiting developmental delay and immunodeficiency. Mol Cell. 2001;8:1175–85.

    Article  PubMed  Google Scholar 

  71. Rossi DJ, Seita J, Czechowicz A, Bhattacharya D, Bryder D, Weissman IL. Hematopoietic stem cell quiescence attenuates DNA damage response and permits DNA damage accumulation during aging. Cell Cycle (Georgetown. Tex). 2007;6:2371–6.

    Article  CAS  Google Scholar 

  72. Qing Y, Wang Z, Bunting KD, Gerson SL. Bcl2 overexpression rescues the hematopoietic stem cell defects in Ku70-deficient mice by restoration of quiescence. Blood. 2014;123:1002–11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. Gutschner T, Haemmerle M, Genovese G, Draetta GF, Chin L. Post-translational regulation of Cas9 during G1 enhances homology-directed repair. Cell Rep. 2016;14:1555–66.

    Article  CAS  PubMed  Google Scholar 

  74. Jayavaradhan R, Pillis DM, Goodman M, Zhang F, Zhang Y, Andreassen PR, et al. CRISPR-Cas9 fusion to dominant-negative 53BP1 enhances HDR and inhibits NHEJ specifically at Cas9 target sites. Nat Commun. 2019;10:2866.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  75. Semenova E, Jore MM, Datsenko KA, Semenova A, Westra ER, Wanner B, et al. Interference by clustered regularly interspaced short palindromic repeat (CRISPR) RNA is governed by a seed sequence. Proc Natl Acad Sci USA. 2011;108:10098–103.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  76. Ran FA, Hsu PD, Lin CY, Gootenberg JS, Konermann S, Trevino AE, et al. Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity. Cell. 2013;154:1380–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  77. Kim S, Kim D, Cho SW, Kim J, Kim JS. Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins. Genome Res. 2014;24:1012–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  78. Guilinger JP, Thompson DB, Liu DR. Fusion of catalytically inactive Cas9 to FokI nuclease improves the specificity of genome modification. Nat Biotechnol. 2014;32:577–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  79. Fu Y, Sander JD, Reyon D, Cascio VM, Joung JK. Improving CRISPR-Cas nuclease specificity using truncated guide RNAs. Nat Biotechnol. 2014;32:279–84.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  80. Slaymaker IM, Gao L, Zetsche B, Scott DA, Yan WX, Zhang F. Rationally engineered Cas9 nucleases with improved specificity. Science. 2016;351:84–8.

    Article  CAS  PubMed  Google Scholar 

  81. Kim D, Kim S, Kim S, Park J, Kim JS. Genome-wide target specificities of CRISPR-Cas9 nucleases revealed by multiplex digenome-seq. Genome Res. 2016;26:406–15.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  82. Zuo E, Sun Y, Wei W, Yuan T, Ying W, Sun H, et al. Cytosine base editor generates substantial off-target single-nucleotide variants in mouse embryos. Science. 2019;364:289–92.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  83. Wienert B, Wyman SK, Richardson CD, Yeh CD, Akcakaya P, Porritt MJ, et al. Unbiased detection of CRISPR off-targets in vivo using DISCOVER-Seq. Science. 2019;364:286–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  84. Strohkendl I, Saifuddin FA, Rybarski JR, Finkelstein IJ, Russell R. Kinetic basis for DNA target specificity of CRISPR-Cas12a. Mol Cell. 2018;71:816–24.e3.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  85. Xu S, Luk K, Yao Q, Shen AH, Zeng J, Wu Y, et al. Editing aberrant splice sites efficiently restores β-globin expression in β-thalassemia. Blood. 2019;133:2255–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Natural Science Foundation of China (grant number 81971886), the Zhujiang Talent Program (grant number 2019QN01Y279) and the Basic and Applied Basic Research Fund of Guangdong Province (grant number 2020A1515010468).

Author information

Authors and Affiliations

  1. Zhanjiang Institute of Clinical Medicine, Zhanjiang Central Hospital, Guangdong Medical University, Zhanjiang, PR China

    Yan Chen, Zhigang Yang & Zhanghui Chen

  2. Department of Hematology, Zhanjiang Central Hospital, Guangdong Medical University, Zhanjiang, PR China

    Ruiting Wen & Zhigang Yang

Authors
  1. Yan Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ruiting Wen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhigang Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhanghui Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhanghui Chen.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, Y., Wen, R., Yang, Z. et al. Genome editing using CRISPR/Cas9 to treat hereditary hematological disorders. Gene Ther 29, 207–216 (2022). https://doi.org/10.1038/s41434-021-00247-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41434-021-00247-9

This article is cited by

Search

Advanced search

Quick links