Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system

• DOI: 10.1038/srep13878
• Publication Date: 2015-09-10T08:58:26Z
• Authors: Xiaolong Wang; Honghao Yu; Anmin Lei; Jiankui Zhou; Wenxian Zeng; Haijing Zhu; Zhiming Dong; Yiyuan Niu; Bingbo Shi; Bei Cai; Jinwang Liu; Shuai Huang; Hailong Yan; Xiaoe Zhao; Guangxian Zhou; Xiaoling He; Xiaoxu Chen; Yuxin Yang; Yu Jiang; Lei Shi; Xiue Tian; Yongjun Wang; Baohua Ma; Xingxu Huang; Lei Qu; Yulin Chen
• Keywords: Gene targeting

Abstract Recent advances in the study of CRISPR/Cas9 system have provided a precise and versatile approach for genome editing various species. However, applicability efficiency this method large animal models, such as goat, not been extensively studied. Here, by co-injection one-cell stage embryos with Cas9 mRNA sgRNAs targeting two functional genes ( MSTN FGF5 ), we successfully produced gene-modified goats either one or both disrupted. The cultured primary fibroblasts was high 60%, while disrupting 98 tested animals 15% 21% respectively 10% double gene modifications. on- off-target mutations target fibroblasts, well somatic tissues testis founder dead animals, were carefully analyzed. results showed that simultaneous several sites achieved demonstrating has potential to become robust efficient engineering tool farm therefore will be critically important applicable breeding.