Table 2 Therapeutic HDR gene-editing strategy for genetic disorders using ZFNs, TALENs and Cas9
From: Homology-directed gene-editing approaches for hematopoietic stem and progenitor cell gene therapy
Disease | Nuclease | Target locus | HDR-gene-editing strategy | Delivery route | Experimental model | References |
|---|---|---|---|---|---|---|
Sickle cell disease & β-thalassemia | ZFNs | HBB | ZFN mRNA and IDLV/ssODN | Electroporation (Harvard apparatus) | HSPCs from healthy donor and SCD patient | [10] |
Cas9 | HBB | Cas9 RNP and plasmid donor | Electroporation (Lonza Nucleofector 4-D) | HSPCs from healthy donor and SCD patient | [23] | |
Cas9 mRNA and ssODN | Electroporation (Neon transfection system and Lonza 4D Nucleofector) | HSPCs from healthy donor | [24] | |||
Cas9 mRNA and IDLV | Electroporation (ECM 830 Square wave electroporator) | HSPCs from SCD patient | [53] | |||
Cas9 RNP and ssODN | Electroporation (Lonza 4D Nucleofector) | HSPCs from healthy donor | [22] | |||
Cas9 RNP and ssODN | Electroporation (Lonza 4D Nucleofector) | HSPCs from healthy donor | [54] | |||
Cas9 RNP and plasmid donor | Electroporation (Lonza Nucleofector 2b) | HSPCs from SCD patient | [55] | |||
X-SCID | ZFNs | IL2RG | ZFN mRNA and IDLV | Electroporation (Lonza) | HSPCs from healthy donor and SCID-X1 patient | [57] |
ZFN and IDLV | Transduction | HSPCs from healthy donor | [58] | |||
Cas9 | Cas9 RNP and AAV6 | Electroporation (Lonza nucleofector 4D) | HSPCs from SCID-X1 patient | [30] | ||
X-linked chronic granulomatous disease | Cas9 | CYBB | Cas9 RNP and ssODN | Electroporation (MaxCyte systems) | HSPCs from X-CGD patient | [29] |
Wiskott–Aldrich syndrome | Cas9 | WAS | Cas9 RNP and AAV6 | Electroporation/transduction (MaxCyte CTX Flow electroporator) | HSPCs from healthy donor and WAS patient | [17] |
Mucopolysaccharidosis | Cas9 | CCR5 | Cas9 RNP and AAV6 | Electroporation/transduction (Lonza nucleofector 4D) | HSPCs from healthy donor | [34] |
Gaucher disease | Cas9 | CCR5 | Cas9 RNP and AAV6 | Electroporation/transduction (Lonza 4D nucleofector) | HSPCs from healthy donor | [59] |
Pyruvate Kinase Deficiency | TALEN& Cas9 | PKLR | TALEN plasmid/Cas9 RNP and plasmid donor | Electroporation (Lonza AMXA II nucleofection system) | HSPCs from healthy donor | [27] |
IPEX syndrome | Cas9 | FOXP3 | Cas9 RNP and AAV6 | Nucleofection/transduction (Lonza 4D nucleofector) | HSPCs from healthy donor | [32] |