动物造模 z-bio 2022-05-17 341

　　Objective To construct lymphocyte activation gene ￣3 (Lag￣3) knockout mice, and to preliminarily analyze the effect of Lag￣3 gene knockout on the phenotype of mice, so as to provide an animal model for subsequent in vivo functional studies of Lag￣3.

　　Methods The knockout mice were constructed by CRISPR/Cas9 technology combined with fertilized egg microinjection, and the knockout-positive mice were screened by molecular identification. ￣3 gene knockout effect, by establishing ConA-induced liver injury model to study the function of Lag￣3 gene in vivo.

　　Results The results of PCR and product sequencing showed that exon2-deficient Lag￣3 knockout mice were obtained. Only the background expression signal of Lag-3 mRNA could be detected in the heart, spleen, lung, macrophages and lymph nodes, and only a very low number of Lag-3 positive cells could be detected in mouse macrophages, spleen cells and lymph nodes. Type analysis showed that the deletion of Lag~3 gene significantly reduced the number of CD3+ T cells in the peripheral blood, bone marrow and spleen of mice induced by ConA, and alleviated the acute liver injury induced by ConA.

　　Conclusion The Lag￣3 gene knockout mouse model was successfully constructed. The in vivo function study during liver injury showed that Lag￣3 deletion can significantly alleviate the occurrence of ConA-induced liver injury.