关键词: 肝豆状核变性, CRISPR/Cas9, 点突变R778L, 小鼠模型

Abstract: Objective R780L mutant mice directed against the human hepatolenticular degeneration R778L mutation type were constructed using a CRISPR/Cas9-mediated gene editing system. Methods The conserved nature of the ATP7B gene and protein in humans and mice was confirmed by BLAST. Microinjection of mouse fertilized eggs was carried out by the CRISPR/Cas9 system, and pathological and physiological tests were performed on the symptoms of hepatolenticular degeneration mouse model. Results The sequences of ATP7B gene and protein between humans and mouse were highly conserved. The homozygous R780L mutant mouse were successfully obtained by CRISPR/Cas9 system, which showed significant liver copper ion deposition and elevated serum ALT and AST and no detectable off-target effects were observed. Conclusion Successfully constructed a R780L mutant mouse model for human hepatolenticular degeneration R778L type using CRISPR/Cas9-mediated gene knock-in system.

Key words: Wilson's disease, CRISPR/Cas9, Point mutation R778L, Mouse model