动物造模,CRISPR-Cas9 z-bio 2022-11-08 341

　　Objective: To obtain albino C57BL/6N mice and expand their application in skin transplantation and embryonic stem cell research.

　　Methods: Cas9mRNA and a series of sgRNAs (single guide RNAs) were designed and synthesized in vitro and injected into the fertilized eggs of mice to destroy the first tyrosinase (TYR) gene sequence necessary for the synthesis of C57BL/6N mouse melanin production. And the second exon, gene mutation was generated, and F0 albino C57BL/6N mice were obtained, and then through repeated backcrossing and selfing, an inbred line of albino C57BL/6N mice was formed.

　　RESULTS: After injection of two pairs of sgRNA, F0 generation albino mice were successfully obtained, and deletion mutations occurred in the first and second exons of the Tyr gene. Homozygous white C57BL/6N mice were generated in their offspring, and finally the mutation type of albino mice was analyzed.

　　Conclusion: The mouse Tyr gene was destroyed by CRISPR-Cas9 technology, and the albino C57BL/6N mouse inbred line was successfully established, which provided a new research tool for future chimera production and tissue transplantation.