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Design of Cas9/sgRNA To construct the recombinant vector for the preparation of sgRNA by in vitro transcription, the two complementary DNA oligos shown in were annealed to be double-stranded, and then sub-cloned into the pUC57-T7-gRNA vector as described. The animals were regularly maintained in the Shaanbei Cashmere Goat Farm of Yulin University. Considering the off-target effects are site-dependent, we believe the off-target mutagenesis can be avoided by choosing a single or several sites. All the protocols involving the use of animals were in accordance with approved guidelines of the Animal Care and Use Committee of the Northwest A&F University (Approval ID: 2011ZX08008-002). Ethical statement Eleven rams (2–3 years old, body weight: 30–50 kg) and 216 ewes (79 donors and 137 recipients, 2–3 years old, body weight: 24–40 kg) were used in this study. Aktivator odin versii 137 2.