动物造模,高酪氨酸血症III型巴马小型猪模型 z-bio 2022-11-03 247

　　Objective: Using CRISPR/Cas9 technology to knock out 4-hydroxyphenylpyruvate oxidase in Bama miniature pigs, to prepare the type III Bama miniature pig model of hypertyrosinemia

　　Methods: The templates for in vitro transcription of sgRNA Hpd and Cas9 were amplified by PCR from plasmids pGL3-U6-gRNA PGKpuromycin and pST1374-NLS flag linker Cas9 respectively, then Cas9 mRNA and sgRNA Hpd were transcribed in vitro, and finally Cas9 mRNA and sgRNA Hpd were co injected into the cytoplasm of fertilized eggs of Bama miniature pigs to prepare the Bama miniature pigs with Hpd gene knockout

　　Results: After intracytoplasmic co injection, 20 fertilized eggs were transplanted to 2 surrogate sows. A total of 4 offspring Hpd gene knockout Bama miniature pigs were obtained

　　Conclusion: This study has successfully prepared a type III Bama miniature pig model of high tyrosinemia, which will provide a better model for the study of tyrosine metabolic pathway