Table 2.
Studies of the applications of CRISPR/Cas9-mediated genome editing in ALS.
| Target | Point mutation | Organism/cell line | Delivery | Results | References |
|---|---|---|---|---|---|
| SOD1 | A272C | Human ALS patient-derived iPSCs | Electroporation | Discovered early biomarkers and pathways dysregulated in ALS | (Wang et al., 2017) |
| L144FVX | Human ALS patient-derived iPSCs | Adenoassociated Virus Vector | Over half of the iPSCs targeted the Src/c-Abl signaling pathway, suggesting that Src/c-Abl may be a potentially useful target for developing new drugs to treat ALS | (Imamura et al., 2017) | |
| E100G | Human ALS patient-derived iPSCs | Nucleofection | Identified activated ERK and JNK signaling are critical drivers of neurodegeneration in mutant SOD1 motor neurons | (Bhinge et al., 2017) | |
| G93A | SOD1-G93A transgenic mouse | Adenoassociated Virus Vector | Disruption of mutant SOD1 enhances the survival of spinal cord motor neurons and improves motor function and life span | (Gaj et al., 2017) | |
| G93A | SOD1-G93A transgenic mouse | Use an AAV vector and inject it into the lumbar subarachnoid space | Deleting mutant SOD1 via CRISPR/Cas9 prolongs survival in an ALS mouse model | (Duan et al., 2020) | |
| G93A | SOD1-G93A transgenic mouse | Intrathecal injection | The mouse treated by split-intein CRISPR base editor had a reduced rate of muscle atrophy, improved neuromuscular function, and up to 40% fewer SOD1 immunoreactive inclusions | (Lim et al., 2020) | |
| G93A | Human SOD1-G93A missense mutation iPSCs | Sendai virus-based vector | The mutant motor neurons accumulated misfolded and aggregated forms of SOD1 in cell bodies, causing axonopathy and aberrant neurotransmission | (Kim et al., 2020) | |
| G93A | SOD1-G93A transgenic mouse | Intracerebroventricular injection | Rescues motor function deficits and extends survival in a SOD1-ALS mouse model | (Chen et al., 2022) | |
| C9ORF72 | Deleted GGGGCC | Human ALS patient-derived iPSCs | Cloned into px300 plasmid and introduced to the iPSCs by nucleofection | Provides a proof-of-principle for using CRISPR-Cas9-mediated excision of the pathogenic C9orf72 repeat expansion as a therapeutic strategy in ALS | (Lopez-Gonzalez et al., 2016) |
| Deleted GGGGCC | Human ALS patient-derived iPSCs | Nucleofection | The mutations lead to increased Ca2+-permeable and enhance selective MNs’ vulnerability to excitotoxicity | (Selvaraj et al., 2018) | |
| Deleted GGGGCC | C9ORF72 patient-derived iPSCs | Sendai virus-based vector | Performed an extensive phenotypic characterization of ALS-iPSCs-derived MNs | (Dafinca et al., 2016) | |
| Deleted GGGGCC | C9ORF72 patient-derived iPSCs | Nucleofection | Proved partial inhibition of an overactivated DNA repair pathway suppresses a cell death pathway is the pathology of ALS | (Lopez-Gonzalez et al., 2019) | |
| Deleted GGGGCC | C9ORF72 patient-derived iPSCs | Adenoassociated Virus Vector 9 | The CRISPR/Cas9-mediated genome correction reduced RNA foci, poly-dipeptides, and haploinsufficiency, major hallmarks of ALS | (Meijboom et al., 2022) | |
| FUS | R521H | FUS patient-derived iPSCs | Nucleofection | Mutations in FUS result in apparent defects in MNs derived from FUS-ALS patients | (Guo et al., 2017) |
| G1566A | FUS patient-derived iPSCs | Electroporation | Discovered early biomarkers and pathways dysregulated in ALS | (Wang et al., 2017) | |
| H517Q | Human ALS patient-derived iPSCs | Nucleofection | Identified activated ERK and JNK signaling as critical drivers of neurodegeneration in mutant FUS MNs | (Bhinge et al., 2017) | |
| R521H, P525L | Human ALS patient-derived iPSCs | Nucleofection | Uncovered a pathway of defective DNA ligation in FUS-linked ALS | (Wang et al., 2018) | |
| R521H, P525L | Human ALS patient-derived iPSCs | Nucleofection | Metabolic dysfunction is not the underlying cause of the ALS-related phenotypes previously observed in these MNs | (Vandoorne et al., 2019) | |
| R524S, P525L | C. legan knockin models | Nucleofection | Autophagy dysfunction likely contributes to protein homeostasis and neuromuscular defects in ALS FUS knockin animals | (Baskoylu et al., 2022) | |
| TARDBP | M337V | TARDBP-M337V patient-derived iPSCs | Nucleofection | The abnormal function of BDNF may explain the aberrant TDP-43 activity | (Tann et al., 2019) |
| M337V | TARDBP-M337V patient-derived iPSCs | Sendai virus-based vector | The MNs with TARDBP mutations impaired mitochondrial Ca2+ uptake contributes to glutamate excitotoxicity | (Dafinca et al., 2020) |
ALS, amyotrophic lateral sclerosis; MNs, motor neurons; iPSCs, induced pluripotent stem cells.