Table 1.
Genome editing of mammalian embryonic stem cells
| Subject | Targeted Gene | Efficiency of Modification * | Off-target Mutation | Genome Editing | Delivery Method | Ref. |
|---|---|---|---|---|---|---|
| NHEJ for InDel | ||||||
| Rat haploid ESCs | Tet1, Tet2, Tet3 | 91% (Single locus), 45% (Double), 14% (Triple) | N.D. | Cas9 | Plasmid/sgRNA | [53] |
| Mouse ESCs | Tet1, Tet2, Tet3, Sry, Uty | 56 ~ 77% (Single; Tet1,2,3), 21% (Triple; Tet1,2,3) | No | Cas9 | Plasmid | [43] |
| HDR for Gene Addition | ||||||
| Human ESCs | GFP into CCR5 | 5.3% | Yes | ZFNs | Lentivirus | [49] |
| Human ESCs | PGK-HygroR into, PIG-A | 0.19 ~ 83.49% | N.D. | ZFNs | Plasmid | [52] |
| Human ESCs | GFP (or PURO) into, AAVS1, OCT4, PITX | 2.9 ~ 15.2% (AAVS1), 0% (OCT4, PITX3) | No (AAVS1, OCT4), Yes (PITX3) | ZFNs | Plasmid | [50] |
| Human ESCs | GFP (and/or PURO), into. AAVS1, OCT4, PITX3 | 9 ~ 22% (AAVS1), 1 ~ 2% (PITX3), 0% (OCT4) | Yes (AAVS1, OCT4), N.D. (PITX3) | TALENs | Plasmid | [51] |
| HDR for Gene Correction or Modification | ||||||
| Human ESCs | Removal of Integrated GFP | 0.24% | N.D. | ZFNs | Plasmid | [52] |
| Human ESCs | Introduction of HindIII cleavage site into EMX1 | 0.4% | No | Cas9 nickase mutant | Plasmid/sgRNA/Oligo | [42] |
| Mouse ESCs | Crytg with 1 bp deletion in exon3 | 0 ~ 44.4% | Yes | Cas9 | Plasmid | [35] |
ESCs; embryonic stem cells, NHEJ; non-homologous end-joining, HDR; homology-directed repair, GFP; green fluorescent protein, PURO; puromycin, sgRNA; single guide RNA, N.D.; not determined.
*Biallelic modification.