Abstract
Beta-thalassemia (β-thalassemia) is an autosomal recessive disorder caused by point mutations, insertions, and deletions in the HBB gene cluster, resulting in the underproduction of β-globin chains. The most severe type may demonstrate complications including massive hepatosplenomegaly, bone deformities, and severe growth retardation in children. Treatments for β-thalassemia include blood transfusion, splenectomy, and allogeneic hematopoietic stem cell transplantation (HSCT). However, long-term blood transfusions require regular iron removal therapy. For allogeneic HSCT, human lymphocyte antigen (HLA)-matched donors are rarely available, and acute graft-versus-host disease (GVHD) may occur after the transplantation. Thus, these conventional treatments are facing significant challenges. In recent years, with the advent and advancement of CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) gene editing technology, precise genome editing has achieved encouraging successes in basic and clinical studies for treating various genetic disorders, including β-thalassemia. Target gene-edited autogeneic HSCT helps patients avoid graft rejection and GVHD, making it a promising curative therapy for transfusion-dependent β-thalassemia (TDT). In this review, we introduce the development and mechanisms of CRISPR/Cas9. Recent advances on feasible strategies of CRISPR/Cas9 targeting three globin genes (HBB, HBG, and HBA) and targeting cell selections for β-thalassemia therapy are highlighted. Current CRISPR-based clinical trials in the treatment of β-thalassemia are summarized, which are focused on γ-globin reactivation and fetal hemoglobin reproduction in hematopoietic stem cells. Lastly, the applications of other promising CRISPR-based technologies, such as base editing and prime editing, in treating β-thalassemia and the limitations of the CRISPR/Cas system in therapeutic applications are discussed.
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Data sharing is not applicable to this article as no new data were created or analyzed in this study.
Abbreviations
- AAV6:
-
Adeno-associated virus 6
- ABE:
-
Adenine base editor
- AsCas12a:
-
Acidaminococcus sp.Cas12a
- AYBE:
-
Adenine transversion base editor
- BE:
-
Base editor
- BER:
-
Base excision repair
- Cas9:
-
CRISPR-associated protein 9
- CBE:
-
Cytosine base editor
- ChiCTR:
-
Chinese Clinical Trial Registry
- CRISPR:
-
Clustered regularly interspaced short palindromic repeats
- crRNA:
-
CRISPR RNA
- DNMT1:
-
DNA methyltransferase 1
- DSB:
-
Double-strand break
- dsDNA:
-
Double-stranded DNA
- ESCs:
-
Embryonic stem cells
- GBE:
-
Glycosylase base editor
- GVHD:
-
Graft-versus-host disease
- HbA:
-
Adult hemoglobin
- HBA :
-
α-globin gene
- HBB :
-
β-globin gene
- HBD :
-
δ-globin gene
- HbE:
-
Hemoglobin E
- HbF:
-
Fetal hemoglobin
- HBG :
-
γ-globin gene
- HDR:
-
Homology-directed repair
- HDAd:
-
Helper-dependent adenovirus
- HIF1α:
-
Hypoxia-inducible factor 1α
- HLA:
-
Human lymphocyte antigen
- HPFH:
-
Hereditary persistence of fetal hemoglobin
- HSCs:
-
Hematopoietic stem cells
- HSPCs:
-
Hematopoietic stem/progenitor cells
- HSCT:
-
Hematopoietic stem cell transplantation
- HUDEP-2:
-
Human umbilical cord blood-derived erythroid progenitor-2
- Indel:
-
Insertion and deletion
- iPSCs:
-
Induced pluripotent stem cells
- LCR:
-
Locus control region
- MMR:
-
Mismatch repair
- nCas9:
-
Cas9 nickase
- NCT:
-
National clinical trial
- NHEJ:
-
Non-homologous end joining
- NSI:
-
NOD-SCID-IL2rg-/-
- PAM:
-
Protospacer adjacent motif
- PBS:
-
Primer binding site
- pegRNA:
-
Prime editing guide RNA
- PE:
-
Prime editor
- PRR:
-
Potential intergenic repressor region
- PRTM1:
-
Protein arginine methyltransferase 1
- PVC:
-
Photorhabdus virulence cassette
- RBC:
-
Red blood cell
- RBM12:
-
RNA binding motif 12
- RNP:
-
Ribonucleoprotein
- ROS:
-
Reactive oxygen species
- RT:
-
Reverse transcriptase
- SCD:
-
Sickle cell disease
- SCID:
-
Severe combined immunodeficiency
- sgRNA:
-
Single-guide RNA
- shRNA:
-
Short hairpin RNA
- SpCas9:
-
Streptococcus pyogenes Cas9
- SSB:
-
Single-strand break
- SSO:
-
Splice-switch oligonucleotide
- ssODN:
-
Single-strand oligodeoxynucleotide
- TadA:
-
tRNA adenosine deaminase
- TALENs:
-
Transcriptional activator-like effector nucleases
- TDT:
-
Transfusion-dependent thalassemia
- TI:
-
Transfusion independence
- tracrRNA:
-
Trans-activating RNA
- TSS:
-
Transcription start site
- UGI:
-
Uracil glycosylase inhibitor
- UNG:
-
Uracil-DNA glycosylase
- ZFNs:
-
Zinc finger nucleases
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This work was supported by the Leading Scholar Award to PH in Jilin University and the National Natural Science Foundation of China (Grant No. 31771624).
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Zeng, S., Lei, S., Qu, C. et al. CRISPR/Cas-based gene editing in therapeutic strategies for beta-thalassemia. Hum. Genet. 142, 1677–1703 (2023). https://doi.org/10.1007/s00439-023-02610-9
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DOI: https://doi.org/10.1007/s00439-023-02610-9