Table 2 Genome editing of mammalian zygotes
Subject | Targeted Gene | Efficiency in Embryos* | Efficiency in Neonates** | Off-target Mutation | Genome Editing | Delivery Method | Remarks | Ref. |
|---|---|---|---|---|---|---|---|---|
NHEJ for InDel | ||||||||
Monkey, zygotes | Nr0b1,Ppar-γ, Rag1 | 18.2 ~ 40.9%, (Single), 9.1 ~ 27.3% (Double) | - | No | Cas9 | mRNA/sgRNA | A set of twin female monkeys with modified Rag1 and Ppar-γ were born. | [33] |
Monkey, zygotes | MECP2 | - | 9.5% (Rhesus), 3.7% (Cynomolgus) | No | TALENs | Plasmid | Three miscarried rhesus and cynomologus male fetuses had Mecp2 mutations. | [34] |
Monkey, zygotes | MECP2 | - | (2.0%) | N.D. | TALENs | mRNA | A modified male monkey appeared to be mosaic. | [47] |
Bovine, zygotes | LDLR | 3.8% | - | N.D. | TALENs | mRNA | Cytoplasmic injection | [46] |
Porcine, zygotes | RELA | 0.5% | - | N.D. | TALENs | mRNA | Cytoplasmic injection | [46] |
Rat, zygotes | IgM | - | 3.9 ~ 5.5% (mRNA) | No (Plasmid), Yes (mRNA) | TALENs | Plasmid or mRNA | Mosaic mutations occurred. Plasmid; Pronuclear, mRNA; Cytoplasmic injection. | [39] |
Rat, Zygotes | Tet1,Tet2,Tet3 | - | 14.3 ~ 18.8% (Double; Tet1,2), 18.6% (Triple) | Yes (Triple) | Cas9 | mRNA/sgRNA | Mosaic mutations occurred. Cytoplasmic injection | [41] |
Mouse, Zygotes | Tet1,Tet2,Tet3, Sry, Uty | - | 8.0 ~ 17.6% (Single), 14.7 ~ 15.3% (Double; Tet1,2) | No (Tet1,Tet2), N.D. (Tet3) | Cas9 | mRNA/sgRNA | Pronuclear injection | [43] |
Mouse, zygotes | Mecp2 | 80 ~ 100%†| - | No | Cas9, nickase mutant | mRNA/sgRNA | Cytoplasmic injection | [42] |
Mouse, zygotes | Exo1 | 1.4 ~ 6.8% | 0 ~ 10.3% | N.D. | TALENs | mRNA | Pronuclear injection | [48] |
Mouse, zygotes | Fgf10 | - | 14.3 ~ 41.7% | N.D. | Cas9 | mRNA/sgRNA | Cytoplasmic injection | [45] |
Mouse, zygotes | Fgf10 | - | 1.3 ~ 1.5% | N.D. | TALENs | mRNA/sgRNA | Cytoplasmic injection | [45] |
HDR for Gene Addition | ||||||||
Mouse, zygotes | mCherry into Nanog, GFP into Oct4 | - | 1.7% (Nanog), 3.0% (Oct4) | Yes, (Nanog, Oct4) | Cas9 | mRNA/sgRNA/Plasmid | Cytoplasmic or pronuclear injection | [40] |
HDR for Gene Correction or Modification | ||||||||
Mouse, zygotes | Addition of V5 to Sox2, lox to Mecp2 | - | 6.0% (Sox2), 0.8% (Mecp2) | Yes (Mecp2) | Cas9 | mRNA/sgRNA/Oligo | Cytoplasmic or pronuclear injection | [40] |
Mouse, zygotes | Crytg with 1 bp deletion in exon3 | - | 4.4 ~ 5.7% | Yes | Cas9 | mRNA/sgRNA/ Oligo | Cytoplasmic injection | [35] |
Mouse, zygotes | Introduction of a STOP codon into Fah | - | 2.0% (wild type), 2.0% (nickase mutant) | Yes, (mutant < WT) | Cas9 WT and mutant | mRNA/sgRNA/Oligo | Pronuclear injection | [44] |