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Efficient generation of gene-modified pigs via injection of zygote with Cas9/sgRNA.

ABSTRACT: Co-injection of zygotes with Cas9 mRNA and sgRNA has been proven to be an efficient gene-editing strategy for genome modification of different species. Genetic engineering in pigs holds a great promise in biomedical research. By co-injection of one-cell stage embryos with Cas9 mRNA and Npc1l1 sgRNA, we achieved precise Npc1l1 targeting in Chinese Bama miniature pigs at the efficiency as high as 100%. Meanwhile, we carefully analyzed the Npc1l1 sgRNA:Cas9-mediated on- and off-target mutations in various somatic tissues and ovaries, and demonstrated that injection of zygotes with Cas9 mRNA and sgRNA is an efficient and reliable approach for generation of gene-modified pigs.

SUBMITTER: Wang Y

PROVIDER: S-EPMC4317696 | biostudies-literature | 2015 Feb

REPOSITORIES: biostudies-literature

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Efficient generation of gene-modified pigs via injection of zygote with Cas9/sgRNA.

Wang Yong Y Du Yinan Y Shen Bin B Zhou Xiaoyang X Li Jian J Liu Yu Y Wang Jianying J Zhou Jiankui J Hu Bian B Kang Nannan N Gao Jimin J Yu Liqing L Huang Xingxu X Wei Hong H

Scientific reports 20150205

Co-injection of zygotes with Cas9 mRNA and sgRNA has been proven to be an efficient gene-editing strategy for genome modification of different species. Genetic engineering in pigs holds a great promise in biomedical research. By co-injection of one-cell stage embryos with Cas9 mRNA and Npc1l1 sgRNA, we achieved precise Npc1l1 targeting in Chinese Bama miniature pigs at the efficiency as high as 100%. Meanwhile, we carefully analyzed the Npc1l1 sgRNA:Cas9-mediated on- and off-target mutations in ...[more]

PMID: 25653176

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