Objective:To construct three gene editing mice,namely,Nrf2 systemic knockout(Nrf2-KO),cardiomyocyte(Nrf2-KO),cardiomyocyte-specific knockout(Nrf2-Flox),and cardiomyocyte overexpression(Nrf2-CTG)mice,to provide a research tool for exploring the specific mechanism of Nrf2 in cardiovascular metabolic diseases.Methods:Nrf2-KO mice and Nrf2-Flox mice were constructed using CRISPR/Cas9 technology;Nrf2-CTG strain mice were constructed using transgenic technology.DNA was extracted from toe clippings of gene-edited mice at 7-10 days of age,and genotypes were determined by PCR and agarose gel electrophoresis,and sequenced.Results:⑴ The Nrf2-KO F0,F1,and F2 mouse genome expanded samples all had clear and single positive bands at the target band(about 750 bp),while the wild-type mice had positive bands at the target band(about 2 931 bp).Using Nrf2-wt-F1/Nrf2-wt-R1 primers,the wild-type mouse genome in the F2 mouse had a positive band at the target band(about 495 bp),while the Nrf2-KO mouse did not have this target band.The deleted bases in the F0 mouse genome were the bases knocked out by the founder mouse.⑵The Nrf2-Flox F0 and F1 mouse genome expanded samples all had clear and single positive bands at the target band(about 150 bp).Wild-type mice did not have this target band.⑶ The Nrf2-CTG F0 and F1 mouse genome expanded samples all had clear and single positive bands at the target band(about 251 bp),and wild-type mice did not have this target band.Conclusion:Nr2-KO mouse model,Nrf2-Flox mouse model and Nrf2-CTG mouse model were successfully constructed by using CRISPR/Cas9 technology and transgenic technology.
关键词
核因子-红细胞2相关因子2/心肌细胞/CRISPR/Cas9/基因编辑/小鼠
Key words
Nuclear factor-erythroid 2 related factor 2/Cardiomyocyte/CRISPR/Cas9/Gene editing/Mice
维普
万方数据