动物造模,高酪氨酸血症III型 z-bio 2022-08-29 283

　　Objective: To use CRISPR/Cas9 technology to knock out 4-hydroxyphenylpyruvate oxidase in Bama minipig to create a hypertyrosinemia type III Bama minipig model.

　　Methods: sgRNA-Hpd and Cas9 templates for in vitro transcription 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 was transcribed. Co-injection with sgRNA-Hpd into the cytoplasm of fertilized eggs of Bama minipigs to prepare Hpd knockout Bama minipigs.

　　RESULTS: After intracytoplasmic co-injection, a total of 20 fertilized eggs were transplanted into 2 surrogate sows. A total of 4 offspring Hpd knockout Bama minipigs were obtained.

　　Conclusion: In this study, a Bama minipig model of hypertyrosinemia type III was successfully prepared, which will provide a better model for the study of tyrosine metabolic pathways.