TABLE 3.

CRISPR-mediated gene knockin in livestock.

Species	Gene	Purpose of manipulation	Type of KI	Approach	SCNT or MI	KI Animals produced	Mosaicism (%)	References
		Agriculture: improvements in
Sheep	SOCS2	Reproductive traits	Point mutation	Crispr/Cas9 BE	MI	3/4 (25%)	3/3 (100%)	Zhou et al. (2019)
	BMPR1B	Reproductive traits	Point mutation	Crispr/Cas9	MI	5/21 (23.8%)	Not stated	Zhou et al. (2018)
Goat	Tβ4	CCR5-targeted KI, cashmere yield	Gene insertion	Crispr/Cas9	SCNT	1	N/A	Li X. et al. (2019)
	FGF5	Cashmere yield	Point mutation	Crispr/Cas9 BE	MI	5/5 (100%)	5/5 (100%	Li G. et al. (2019)
	GDF9	Reproductive traits	Point mutation	Crispr/Cas9	MI	4/17 (23.5%)	2/4 (50.0%)	Niu et al. (2018)
	FAT-1	Disease resistance	Gene insertion	Crispr/Cas9	SCNT	1 from 8 pregnancies	N/A	Zhang J. et al. (2018)
Cattle	Pc	Generation of a polled genotype	Gene insertion	Crispr/Cas12a	SCNT	1, died on D1 after birth	N/A	Schuster et al. (2020)
	NRAMP1	Tuberculosis resistance	Gene insertion	Crispr/Cas9n	SCNT	9	N/A	Gao et al. (2017)
	IARS	Correction of IARS syndrome	Gene insertion	Crispr/Cas9	SCNT	5 viable fetuses	N/A	Ikeda et al. (2017)
Pig	PBD-2	Disease-resistant pigs	Gene insertion	Crispr/Cas9	SCNT	5 pigs	N/A	Huang et al. (2020)
	MSTN	Meat production	Gene insertion	Crispr/Cas9	SCNT	2 pigs	N/A	Zou Y.-L. et al. (2019)
	UCP1	Reproduction traits	Gene insertion	Crispr/Cas9	SCNT	12 piglets	N/A	Zheng et al. (2017)
	MSTN	Meat production	Point mutation	Crispr/Cas9	SCNT	1 stillborn piglet	N/A	Wang K. et al. (2016)
	MSTN	MSTN-KO without selectable marker	Gene insertion	Crispr/Cas9	SCNT	2 piglets	No	Bi et al. (2016)
	RSAD2	Generation of pigs with viral resistance	Gene insertion	Crispr/Cas9	SCNT	1 pig	No	Xie et al. (2020)
		Biomedical applications:
Sheep	ALPL	Model of hypophosphatasia	Point mutation	Crispr/Cas9	MI	6/9 (66.6%)	No	Williams et al. (2018)
	PPT1	Infantile neuronal ceroid lipofuscinoses	Point mutation	Crispr/Cas9	MI	6/24 (25.0%)	Not stated	Eaton et al. (2019)
	tGFP	Rosa26-targeted KI	Gene insertion	Crispr/Cas9	MI	1/8 (12.5%)	Not stated	Wu et al. (2016)
	OTOF	Hearing loss phenotype	Point mutation	Crispr/Cas9	MI	8/73 (11.0%)	2/8 (25.0%)	Menchaca et al. (2020b)
Cattle	CMAH	Xenotransplantation	Point mutation	Crispr/Cas12a	SCNT	2	N/A	Perota et al. (2019)
Pig	hF9	Gene therapy for hemophilia B pigs	Gene insertion	Crispr/Cas9	SCNT	5 pigs	N/A	Chen et al. (2020)
	BgEgXyAp	Salivary gland as bioreactor	Gene insertion	Crispr/Cas9	SCNT	4 piglets (1/4 alive)	N/A	Li G. et al. (2020)
	hIAPP	Type 2 diabetic miniature pig model	Gene insertion	Crispr/Cas9	SCNT	24	N/A	Zou X. et al. (2019)
	SNCA	Parkinson’s disease model	Gene insertion	Crispr/Cas9	SCNT	8 piglets	N/A	Zhu et al. (2018)
	HTT	Huntingtin KI model	Gene insertion	Crispr/Cas9	SCNT	6 piglets	N/A	Yan et al. (2018)
	GGTA1	Xenotransplantation	Gene insertion	FokI-dCas9	SCNT	2 piglets	N/A	Nottle et al. (2017)
	tdTomato	porcine Oct4 reporter system	Gene insertion	Crispr/Cas9	SCNT	2 piglets	N/A	Lai et al. (2016)
	hALB	Tg animals as bioreactors	Gene insertion	Crispr/Cas9	MI	16/16 (100%)	1/16 (6.3%)	Peng et al. (2015)
	GFP	H11-targeted KI	Gene insertion	Crispr/Cas9	SCNT	1 piglet	N/A	Ruan et al. (2015)

SCNT, somatic cell nuclear transfer; MI, zygote microinjection; BE, base editing; N/A, not applicable.