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Int J Biol Sci 2018; 14(4):427-436. doi:10.7150/ijbs.23890 This issue Cite

Research Paper

Generation of Cashmere Goats Carrying an EDAR Gene Mutant Using CRISPR-Cas9-Mediated Genome Editing

Fei Hao1,2, Wei Yan1, Xiaocong Li1, Hui Wang1, Yingmin Wang1, Xiao Hu1, Xu Liu1, Hao Liang1, Dongjun Liu1✉

1. State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, Inner Mongolia University, Hohhot, 010070, P. R. China.
2. Wulanchabu Academy of Agricultural and Animal Husbandry Sciences, Wulanchabu, 012000, P. R. China.

✉ Corresponding author: Dongjun Liu (email: nmliudongjuncom)

Citation:
Hao F, Yan W, Li X, Wang H, Wang Y, Hu X, Liu X, Liang H, Liu D. Generation of Cashmere Goats Carrying an EDAR Gene Mutant Using CRISPR-Cas9-Mediated Genome Editing. Int J Biol Sci 2018; 14(4):427-436. doi:10.7150/ijbs.23890. https://www.ijbs.com/v14p0427.htm
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Abstract

Graphic abstract

In recent years, while the use of the clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR-associated protein 9 (Cas9) (CRISPR-Cas9) system for targeted genome editing has become a research hotspot, it has, to date, not proved adequate for genome editing in large mammals, such as goats. In this study, two opposite single-guide RNAs (sgRNAs) were designed for complete EDAR gene targeting in Cashmere goats, and co-transfected with a plasmid encoding Cas9 into goat fibroblasts. Among the 89 cell lines obtained through the cultivation of clonal cell lines, 62 were positive for EDAR gene targeting. Nine types of mutations were identified by sequencing analysis, and the mutation efficiency was 69.7%. Using one of these cell lines, EDAR gene-targeted Cashmere goat embryos were prepared by somatic cell cloning. Developed embryos were transferred to 79 Cashmere goat recipients, and, after a gestation period of five months six male EDAR gene-targeted Cashmere goats were born. Although only two of these goats survived, they had abnormal primary hair follicles and no hair on the top of their heads, which are the distinctive features of the EDAR gene-targeted Cashmere goats. Thus, this study provides a valuable animal model for future studies on EDAR gene-related phenotypes and hair follicle growth and development and shows that the CRISPR-Cas9 system can be used to edit genes in large mammals.

Keywords: CRISPR-Cas9, EDAR, goat, hair follicle, SCNT

Citation styles

APA
Hao, F., Yan, W., Li, X., Wang, H., Wang, Y., Hu, X., Liu, X., Liang, H., Liu, D. (2018). Generation of Cashmere Goats Carrying an EDAR Gene Mutant Using CRISPR-Cas9-Mediated Genome Editing. International Journal of Biological Sciences, 14(4), 427-436. https://doi.org/10.7150/ijbs.23890.

ACS
Hao, F.; Yan, W.; Li, X.; Wang, H.; Wang, Y.; Hu, X.; Liu, X.; Liang, H.; Liu, D. Generation of Cashmere Goats Carrying an EDAR Gene Mutant Using CRISPR-Cas9-Mediated Genome Editing. Int. J. Biol. Sci. 2018, 14 (4), 427-436. DOI: 10.7150/ijbs.23890.

NLM
Hao F, Yan W, Li X, Wang H, Wang Y, Hu X, Liu X, Liang H, Liu D. Generation of Cashmere Goats Carrying an EDAR Gene Mutant Using CRISPR-Cas9-Mediated Genome Editing. Int J Biol Sci 2018; 14(4):427-436. doi:10.7150/ijbs.23890. https://www.ijbs.com/v14p0427.htm

CSE
Hao F, Yan W, Li X, Wang H, Wang Y, Hu X, Liu X, Liang H, Liu D. 2018. Generation of Cashmere Goats Carrying an EDAR Gene Mutant Using CRISPR-Cas9-Mediated Genome Editing. Int J Biol Sci. 14(4):427-436.

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