Table 1.
Summary of CRISPR-mediated therapeutic strategies to rescue the DMD phenotype
| Subject | Strategy | Nuclease | DMD mutation | Therapeutic target gene region(s) | Model(s) | Delivery | Reference |
|---|---|---|---|---|---|---|---|
| Therapeutic approach | exon reframing | SpCas9 | DMD exon 48–50 deletions DMD nonsense mutation in exon 51 | DMD exon 51 | human DMD myoblasts | electroporation | 37 |
| SpCas9 | DMD exon 45–52 deletions | DMD exon 53 | human DMD myoblasts | adenovirus | 38 | ||
| SpCas9 | DMD exon 44 deletion | DMD exon 45 | human iPSCs | electroporation | 39 | ||
| CjCas9 | Dmd nonsense mutation in exon 23 | Dmd exon 23 | DMD mice | All-in-one AAV9 | 40 | ||
| LbCpf1 | DMD exon 48–50 deletions DMD nonsense mutation in exon 51 | DMD exon 51 | human iPSCs | nucleofection | 41 | ||
| exon deletion | SpCas9 | Dmd nonsense mutation in exon 23 | Dmd intron 22 and 23 | mdx mice | AAV9 | 42 | |
| SpCas9 | DMD exon 46–51 deletions DMD exon 46–47 deletions | DMD intron 44 and 55 | human DMD myoblasts | nucleofection | 43 | ||
| SaCas9 | Dmd nonsense mutation in exon 23 | Dmd intron 22 and 23 | mdx mice | AAV8 | 44 | ||
| SaCas9 | Dmd nonsense mutation in exon 23 | Dmdexon 23 | Ai9 mdx mice | AAV9 | 45 | ||
| exon skipping | SpCas9 | DMD point mutation in intron 47, exon 51 | DMD exon 47A, exon 51 | human iPSCs | nucleofection | 46 | |
| SpCas9 | DMD exon 44 deletion | DMD splice site of exon 43 or exon 45 | human iPSCs | nucleofection | 47 | ||
| SpCas9 | DMD nonsense mutation in exon 53 | DMD splice acceptor site of exon 53 | human DMD myoblasts | adenovirus | 48 | ||
| SpCas9 | Dmd exon 50 deletion | Dmd splice acceptor site of exon 51 | DMD mice | AAV9 | 49 | ||
| SpCas9 | DMD exon 50 deletion | DMD splice acceptor site of exon 51 | canine model of DMD | AAV9 | 50 | ||
| homology-directed repair | SpCas9 | Dmd nonsense mutation in exon 23 | Dmd exon 23 | mdx mice | injection | 51 | |
| SpCas9 | Dmd nonsense mutation in exon 23 | Dmd exon 23 | mouse muscle stem cells | adenovirus | 52 | ||
| SpCas9 | Dmd nonsense mutation in exon 53 | Dmd exon 53 | mdx4cv mice | AAV6 | 53 | ||
| SpCas9 | DMD exon 44 deletion | DMD exon 44 | human iPSCs | electroporation | 39 | ||
| SpCas9 | DMD exon 7 skipping | DMD splice acceptor site of intron 6 and exon 7 boundary | canine model of DMD | injection | 54 | ||
| LbCpf1 | Dmd nonsense mutation in exon 23 | Dmd exon 23 | mdx mice | injection | 41 | ||
| base editing (exon skipping) | TAM based on SaCas9 (Cytosine base editor) | DMD exon 51 deletion | DMD splice site of exon 50 | human iPSCs | lipotransfection | 55 | |
| base editing (correction) | ABE7.10 | Dmd nonsense mutation in exon 20 | Dmd exon 20 | DMD mice | trans-splicing AAV | 56 | |
| transcriptional modulation | CRISPRa (dCas9-VP160) | DMD exon 45–52 deletions | UTRN A, B promoter | immortalized DMD patient muscle cells | Electroporation | 57 | |
| CRISPRa (dCas9-VP160) | Dmd nonsense mutation in exon 23 | Lama1 promoter | mouse myoblasts, mdx/rag mice | transfection, injection, electroporation | 58 | ||
| CRISPRa (dCas9-VP64) | Dmd nonsense mutation in exon 23 | klotho and Utrn | mdx mice | AAV9 | 59 | ||
| SaCas9 | DMD exon 46–51 deletions | 3′ UTR of UTRN inhibitory miRNA target region | human iPSCs | electroporation | 60 | ||
| CRISPRi (dCas9-KRAB) | epigenetic dysregulation of DUX4 | DUX4 promoter or DUX4 exon 1 | human FSHD myocytes | lentivirus | 61 |
AAV, adeno-associated viral vector; ABE, adenine base editor; CRISPRa, CRISPR activator; CRISPRi, CRISPR interference; FSHD, facioscapulohumeral muscular dystrophy; hiPSC, human induced pluripotent stem cell; TAM, targeted AID mediate mutagenesis.