Abstract: Objective The aim of this experiment was to construct p53 apoptosis stimulating protein 2 (ASPP2) gene liver conditional knockout mice by Cre-loxP system. Methods The ASPP2-flox mice were constructed by inserting loxP sequence at both ends of ASPP2 gene with designed sgRNA sequence. The F0 and F1 mice were genotyped by PCR amplification and gene sequencing. The ASPP2-flox F1 mice were mated with Alb-Cre expressing tool mice to obtain liver ASPP2 liver conditional knockout mice, and the genotypes were identified by PCR. The expression of ASPP2 protein in liver tissues of ASPP2 gene knockout mice was detected by Western blot and immunohistochemistry. Results The PCR and sequencing results of F0 and F1 generation mice showed that the ASPP2-flox mice were successfully constructed, and the genotype of F1 generation heterozygotes was ASPP2fl/-; after hybridization with Alb-Cre mice, the F1 generation heterozygotes were backcrossed with ASPP2fl/- mice, and the genotype was ASPP2fL/fLCreT, namely ASPP2 liver conditional knockout mice; the ASPP2 protein expression in liver tissues of ASPP2fL/fLCreT mice by Western blot and immunohistochemistry detection was significantly decreased. Conclusion The ASPP2 gene liver conditional knockout mice is successfully constructed based on Cre-loxP system, which might be beneficial to further experimental study.

Key words: Cre-loxP system, p53 apoptosis stimulating protein 2, Conditional knockout, Mice