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Examples of genome editing-mediated genetic modification in livestock

 
Subject Target Gene Efficiency in zygotes* Efficiency in Live Born Off-target Mutation Gene Editing Delivery Remarks Reference
NHEJ (non-homologous end-joining)
Bovine zygotes MSTN - 19%** N.D. TALEN mRNA Cytoplasmic injection Proudfoot et al., 2015
Bovine zygotes LDLR 3.8% N.D. TALEN mRNA Cytoplasmic injection Carlson et al., 2012
Porcine zygotes MSTN 20% (biallelic) No Cas9 Cas9 protein, sgRNA Mosaicism, electroporation Tanihara et al., 2016
Porcine zygotes GGTA1 50% (biallelic) No Cas9 Plasmid Mosaicism, cytoplasmic injection Petersen et al., 2016
Porcine zygotes CD163 9%** N.D. Cas9 mRNA/sgRNA Cytoplasmic injection Whitworth et al., 2016
Porcine zygotes MITF 5% (biallelic)** No Cas9 mRNA/sgRNA Cytoplasmic injection Wang et al., 2015b
Porcine zygotes Npc1l1 11%** No Cas9 mRNA/sgRNA Mosaicism. cytoplasmic injection Wang et al., 2015c
Porcine zygotes vWF 15%** No Cas9 mRNA/sgRNA Cytoplasmic injection Hai et al., 2014
Porcine zygotes RELA 0.5% N.D. TALEN mRNA Cytoplasmic injection Carlson et al., 2012
Ovine zygotes MSTN, ASIP, BCO2 6% (triple KO) No Cas9 mRNA/sgRNA Cytoplasmic injection Wang et al., 2016
Ovine zygotes MSTN 4.6% 23% (homo-zygous KO) Yes (20% in mutants) Cas9 mRNA/sgRNA Mosaicism, cytoplasmic injection Crispo et al., 2015
Ovine zygotes MSTN 4%** N.D. TALEN mRNA Cytoplasmic injection Proudfoot et al., 2015
Caprine zygotes MSTN, FGF5 13% (double KO) Yes (23% in mutants) Cas9 mRNA/sgRNA Cytoplasmic injection Wang et al., 2015a
Chicken PGCs OVM > 90% G1 from #372: 58% G1 from #376: 48% No Cas9 Plasmid Transfection Oishi et al., 2016
PKFs CMAH, GTA1 0.5% ** (double KO) N.D. Cas9 mRNA Sequential SCNTs of edited cell lines Fischer et al., 2016
HDR (homology-directed repair)
BEFs POLLED introgression 7% (Day 70) No TALEN mRNA, Oligo DNA SCNT of edited cell lines Carlson et al., 2016
GFFs hLF insertion after BLG KO 40% (3 mo) N.D. TALEN mRNA, pBLG-hLF-puro SCNT of edited cell lines Cui et al., 2015
*Genetically modified embryos per injected zygote (%).
**Genetically modified offspring per injected embryo (%).
N.D.: not determined.
PKF: Porcine Kidney Fibroblast.
PGC: Primordial Germ Cell.
GFF: Goat Fetal Fibroblast. Bovine Embryo Fibroblasts.
SCNT: Somatic Cell Nuclear Transfer.