Mutation Screening in Rabbits

The gene-edited rabbits were successfully created in previous research by our group.11, 12, 13, 14 The protocol for mutation detection has been described previously.17 DNA from knockout rabbits was isolated using the TIANamp genomic DNA kit (Tiangen, Beijing, China) according to the manufacturer’s instructions. The PCR products were gel-purified and cloned into pGM-T (Tiangen, Beijing, China). At least 30 positive plasmid clones were sequenced by Comate Bioscience (Changchun, China) with Sanger sequencing and analyzed using commercial software (DNAMAN, Lynnon Biosoft, San Ramon, CA, USA, and the basic local alignment search tool, National Center for Biotechnology Information, Bethesda, MD, USA).