Table 2.
Overview of diseases addressed by gene editing in human primary hematopoietic cells.
| Target Modification | Cell Source/Cell Type | Gene (Targeted Modification) | Gene Editing Strategy | Nuclease Delivery | Repair Donor Template | Efficiency of Gene Editing | Reference |
|---|---|---|---|---|---|---|---|
| β-Hemoglobinopathies | |||||||
| Gene addition/correction | iPSCs | HBB (c.20 A>T) | ZFN | pDNA | pDNA | 50% HDR | [125] |
| iPSCs | HBB (c.20 A>T) | ZFN | pDNA | pDNA | 38% HDR | [126] | |
| iPSCs | HBB (c.316-197C>T, c.126_129delCTTT) | TALEN | pDNA | pDNA | 68% HDR | [127] | |
| iPSCs | HBB: c.-78A>G, c.126_129delCTTT | CRISPR/Cas9 | pDNA | PiggyBac | 23% HDR | [128] | |
| HSCs | HBB (c.20 A>T) | ZFN | mRNA | IDLV/ssODN | 40% HDR | [82] | |
| iPSCs | HBB (c.52A>T) | CRISPR/Cas9 | pDNA | dsDNA | 17% HDR | [129] | |
| iPSCs | HBB (c.316-197C>T) | CRISPR/Cas9 & TALEN | pDNA | PiggyBac | 12 & 33% HDR | [130] | |
| iPSCs | HBB (c.20 A>T) | CRISPR/Cas9 | pDNA | dsDNA | 40% HDR | [131] | |
| Human Embryos | HBB (c.126_129delCTTT) | CRISPR/Cas9 | mRNA | ssODNs | 14% HDR | [107] | |
| HSPCs | HBB (Exon1 & c.20 A>T) | CRISPR/Cas9 | mRNA/RNP | AAV6 | 10% HDR | [106] | |
| iPSCs | HBB (c.126_129delCTTT) | CRISPR/Cas9 | pDNA | ssODNs | 5% HDR | [132] | |
| iPSCs | HBB (c.126_129delCTTT) | CRISPR/Cas9 | pDNA | dsDNA | 57% HDR | [133] | |
| HSPCs | HBB (c.20 A>T) | CRISPR/Cas9 | mRNA | IDLV | 20% HDR | [134] | |
| HSPCs | HBB (c.20 A>T) | CRISPR/Cas9 | RNP | ssODNs | 33% HDR | [81] | |
| iPSCs | HBB (c.126_129delCTTT) | CRISPR/Cas9 | pDNA | dsDNA | NA | [135] | |
| Human Embryos | HBB (c.126_129delCTTT) | CRISPR/Cas9 | RNP | ssODNs | 25% HDR | [136] | |
| HSPCs | HBB (c.20 A>T) | CRISPR/Cas9 | mRNA/RNP | ssODNs | 9% HDR | [137] | |
| HSPCs | HBB (c.126_129delCTTT) | CRISPR/Cas9 | pDNA | ssODNs | 54% HDR | [138] | |
| iPSCs | HBB (Exon1, 3′ UTR, c.52A>T, c316-197C>T & c.126_129delCTTT) | CRISPR/Cas9 | pDNA | pDNA | NA | [139] | |
| Human Embryos | HBB (c.-28A>G) | Base Editor | mRNA | - | 23% BE | [140] | |
| HSPCs | HBB (c.93-21G>A) | CRISPR/Cas9, TALENs & ZFN | mRNA | ssODNs | 8% HDR | [141] | |
| HSPCs | HBB (c.20 A>T) | CRISPR/Cas9 | RNP | AAV6 | 70% HDR | [142] | |
| Gene disruption | HSPCs | BCL11A (GATA 1) | CRISPR/Cas9 | LV | - | NA | [143] |
| HSPCs | HBG1/2 promoter | CRISPR/Cas9 | LV | - | 77% NHEJ | [144] | |
| HSPCs | HBD-HBB | CRISPR/Cas9 (SaCas9) | pDNA | - | 31% NHEJ | [145] | |
| HSPCs | BCL11A (Exon 2) | CRISPR/Cas9 | pDNA/mRNA | - | 13% NHEJ | [146] | |
| HSPCs | BCL11A (Exon 2, GATAA) | ZFN | mRNA | - | 45–50% NHEJ | [147] | |
| HSPCs | HBA MCS-R2 enhancer | CRISPR/Cas9 | dsDNA | - | 60% NHEJ | [148] | |
| HSPCs | HBG-HBD, HBD-HBB | CRISPR/Cas9 | pDNA | - | 20% NHEJ | [149] | |
| HSPCs | HBG1/2 promoter | CRISPR/Cas9 | ADV | - | 24% NHEJ | [150] | |
| HSPCs | BCL11A (Exon 2) | ZFN | mRNA | - | 72% NHEJ | [151] | |
| HSPCs | HBB (c.93-21G>A) | CRISPR/Cas9 & TALEN | RNP and mRNA | - | 90% NHEJ | [152] | |
| HSPCs | BCL11A (GATA 1) | CRISPR/Cas9 | RNP | - | 87% NHEJ | [153] | |
| HSPCs | HBG1/2 | TALEN | mRNA | - | 74% NHEJ | [154] | |
| Severe Combined Immunodeficiencies | |||||||
| Gene addition/correction | T-cells | IL2RG (exon 5) | ZFN | pDNA | pDNA | 7% HDR | [155] |
| HSPCs | IL2RG | ZFN | IDLV | IDLV | 39% HDR | [156] | |
| HSPCs | IL2RG | ZNF | mRNA | IDLV | 6% HDR | [43] | |
| iPSCs | JAK3 (c.1837C>T) | CRISPR/Cas9 | pDNA | pDNA | 73% HDR | [157] | |
| HSPCs | IL2RG (c.691G>A) | CRISPR/Cas9 & ZNF | mRNA | AAV6 | 27% HDR in CD34+CD133+CD90+ | [98] | |
| T-cells | IL2RG (c.800delA/c.530A>G) | CRISPR/Cas9 | RNP | ssDNA/dsDNA | 25/22% HDR | [158] | |
| HSPCs | IL2RG | CRISPR/Cas9 | RNP | AAV | 45% HDR | [159] | |
| Wiskott–Aldrich Syndrome | |||||||
| Gene addition/correction | iPSCs | WAS | ZNF | pDNA | pDNA | NA | [160] |
| HSPCs | WAS | CRISPR/Cas9 | RNP | AAV6 | 60% HDR | [161] | |
| Fabry Disease | |||||||
| Gene addition/correction | HSPCs | GLA (TI in α-globin 5′ UTR) | AAV6 | RNP | AAV6 | NS (16x G:LA expression) | [162] |
| Hurler Syndrome | |||||||
| Gene addition/correction | HSPCs | IDUA (TI in α-globin 5′ UTR) | AAV6 | RNP | AAV6 | NS (171x IDUA expression) | [162] |
| Wolman Disease | |||||||
| Gene addition/correction | HSPCs | LAL (TI in α-globin 5′ UTR) | AAV6 | RNP | AAV6 | 0.7 TI LAL copies/cell | [162] |
| Chronic Granulomatous Disease | |||||||
| Gene addition/correction | iPSCs | AAVS1 | TALENs | pDNA | pDNA | 50% HDR | [163] |
| iPSCs | CYBB (Int. 1 T>G) | ZFN | mRNA | AAV6 | 57% HDR | [164] | |
| iPSCs | AAVS1 | ZFN | mRNA | pDNA | 80% HDR | [165] | |
| HSPCs | AAVS1 | CRISPR/Cas9 | LV | AAV6 | 67% HDR | [97] | |
| iPSCs | CYBB | CRISPR/Cas9 | pDNA | pDNA | 17% HDR | [166] | |
| HSPCs | CYBB (C676T) | CRISPR/Cas9 | mRNA | ssODN | 21% HDR | [167] | |
| iPSCs | NCF1B, NCF1C | CRISPR/Cas9 | mRNA | pDNA/rAAV2 | 90% HDR | [168] | |
| iPSCs | NCF1 | CRISPR/Cas9 | pDNA | pDNA | 43–47% | [169] | |
| HSPCs | CYBB | CRISPR/Cas9 | mRNA | ssODN | 80% | [89] | |
| Immunodysregulation Polyendocrinopathy Enteropathy X-Linked | |||||||
| Gene addition/correction | HSPCs | FOXP3 | CRISPR/Cas9 | RNP | rAAV6 | 29% HDR | [170] |
| Hyper IgM Syndrome | |||||||
| Gene addition/correction | T-cells | CD40L | TALEN | mRNA | rAAV | 36–47% | [171] |
| T-cells, CD34+ | CD40L cDNA | TALEN & CRISPR/Cas9 | mRNA | IDLV/AAV6 | 31- 34% | [99] | |
| Fanconi Anemia | |||||||
| Gene addition/correction | iPSCs | FANCA | ZFN | ADV | IDLV | 40% HDR | [172] |
| HSPCs | FANCD1 (Exon 8) | CRISPR/Cas9 | Plasmid | ssDNA | NA | [173] | |
| HSPCs | FANCA | ZFN | mRNA | IDLV | 14% HDR | [174] | |
| HSPCs | (c.3558insG, c.295C>T), FANCB, FANCC (c.67delC), FANCD1/BRACA2 (c.1596delA), FANCD2 (c.718delT) | CRISPR/Cas9 | pDNA | - | NHEJ | [175] | |
| Hemophilia A | |||||||
| Gene addition/correction | iPSC | F8 (Inv 1) | TALENs | pDNA | - | Inversion 1% | [176] |
| iPSC | F8 (Inv 1 & 22) | CRISPR/Cas9 | pDNA | - | Inversion 7% | [177] | |
| iPSC | F8 (Inv 22) | TALENs | pDNA | pDNA | 63% HDR | [178] | |
| iPSC | F8 | CRISPR/Cas9 | RNP | pDNA | 66% HDR | [179] | |
| Hemophilia B | |||||||
| Gene addition/correction | Germline cells | F9 (exon 8) | CRISPR/Cas9 | RNP | ssDNA | 53% HDR | [180] |
| HSPC | F9 (TI in α-globin 5′ UTR) | CRISPR/Cas9 | AAV6 | RNP | 1 TI F9 copy/cell | [162] | |
| Amegakaryocytic Thrombocytopenia | |||||||
| Gene correction | HSPCs | MPL (c.814T>C) | CRISPR/Cas9 | RNP | ssODN | NA | [181] |
| HIV AIDS | |||||||
| Gene disruption | Th cells | CCR5 | TALEN | mRNA GMP electroporation | - | >60% cells, 40% biallelic | [182] |
| HSPC | CCR5 | CRISPR/Cas9 | pDNA electroporation | - | 27% | [183] | |
| Gene silencing | T cells | CCR5 & CXCR4 | TALE epigenome modifier | mRNA electroporation | - | % CpG methylation (10–90% CCR5, 2–13% CXCR4) | [184] |
Abbreviations: AAV—adeno-associated virus vectors; ADV—Adenovirus; HDR—homology-directed repair; IDLV—integrase-deficient; iPSCs—induced pluripotent stem cells; LCLs—lymphoblastic cell lines; LV—lentiviral vector; NA—Not Available; NS—not specified; NHEJ—non-homologous end joining; pDNA—plasmid DNA; RNP—ribonucleoprotein; ssDNA—Single-stranded DNA; ssODN—single-stranded donor oligonucleotides; TI—targeted integration.