Generation and phenotypic analysis of TD02 knockout mice / 中国药理学通报

ABSTRACT

Aim To construct TD02 gene knockout C57BL/6 mice, and meanwhile to study their phenotypes preliminarily. Methods The sgRNA plasmids were designed and constructed according to the principle of CRISPR-Cas9 technology, and were microinjected into mouse zygotes after transcription in vitro with Cas9 plasmids. FO generation mice with multiple genotypes were obtained by embryo transplantation. The PCR method was used to determine the genotype, and the positive F0 generation mice were obtained. F1 generation heterozygous mice were obtained by backcrossing the positive F0 generation mice and the wild type mice. The positive F1 generation heterozygous mice were matched to each other to obtain F2 generation homozygous mice. And their growth characteristics, reproductive ability, and progeny survival rate were observed and analyzed respectively. TDO2 protein expression in liver tissues of homozygous TDO2 gene knockout mice was detected by Western blot. Results TDO2 gene knockout mice were successfully bred and identified, and the genotypes of offspring mice were successfully identified by PCR. The results showed that TDO2 gene knockout was successfully conducted and TDO2 protein expression was not detected in liver tissues of TDO2 gene knockout mice. No abnormal changes were observed in diet, body weight, breeding ability of TDO2 knockout mice. Conclusions TDO2 gene knockout mice have been initially established, which would provide experimental means for studying the biological function of TDO2 gene and its regulatory role in diseases.