1. Efficient generation of gene-modified pigs via injection of zygote with Cas9/sgRNA.
- Author
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Wang Y, Du Y, Shen B, Zhou X, Li J, Liu Y, Wang J, Zhou J, Hu B, Kang N, Gao J, Yu L, Huang X, and Wei H
- Subjects
- Alleles, Animals, Base Sequence, Female, Gene Targeting methods, Membrane Proteins chemistry, Membrane Proteins genetics, Molecular Sequence Data, Mosaicism, Organ Specificity genetics, Ovary metabolism, RNA Editing, RNA, Guide, CRISPR-Cas Systems, Animals, Genetically Modified, CRISPR-Cas Systems genetics, Microinjections methods, RNA, Messenger, Swine, Zygote metabolism
- 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.
- Published
- 2015
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