高酪氨酸血症III型巴马小型猪模型 z-bo 2020-12-29 306

　　Purpose: Use CRISPR/Cas9 technology to knock out 4-OH phenylpyruvate dioxygenase (4HPPD/HPD) in Bama miniature pigs to prepare a model of hypertyrosinemia type III Bama miniature pigs.

　　Method: The sgRNA-Hpd and Cas9 in vitro transcription templates were amplified by PCR from the plasmids pGL3-U6-gRNA-PGKpuromycin and pST1374-NLS-flag-linker-Cas9 respectively, and then Cas9mRNA and sgRNA-Hpd were transcribed in vitro, and finally Cas9mRNA Co-injected with sgRNA-Hpd into the cytoplasm of fertilized Bama minipigs to prepare Hpd knockout Bama minipigs.

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

　　Conclusion: This study has successfully prepared a hypertyrosinemia type III Bama minipig model, which will provide a better model for the study of tyrosine metabolism pathways.