Construction of GPR183 knockout mice and its function in acute liver injury

• Main Authors: WU Xuewen, YIN Siqi, QIAN Ying, DU Bing, QIN Juliang, CHEN Jinlian
• Format: Article
• Language: zho
• Published: Editorial Office of Journal of Third Military Medical University 2021-09-01
• Series: Di-san junyi daxue xuebao
• Subjects: crispr/cas9 gene editing technology; g protein coupled receptor; gene knockout; acute liver injury
• Online Access: http://aammt.tmmu.edu.cn/Upload/rhtml/202103181.htm

Objective To construct a GPR183 knockout mouse model by genome editing using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system, and to analyze the role of GPR183 receptor in lymphocyte regulation and acute liver injury. Methods Firstly, GPR183 full-gene knockout homozygous F2 generation mice were constructed using CRISPR/Cas9 gene editing technique. Then, the expression level of GPR183 protein in the liver and spleen tissues of wild-type (WT) and knockout (KO) mice was detected by Western blotting and the Results were compared; the distribution of spleen lymphocytes and the expression of IL-17 and IL-22 in WT and KO mice were analyzed by immunofluorescence (IF) assay. Finally, the function of GPR183 knockout was explored in mouse model with acute liver injury. Results GPR183 KO mice could grow and reproduce normally, and F2 homozygous KO mice had partial deletion of gene fragments as compared to WT mice. Meanwhile, Western blotting showed that GPR183 expression was missing in the liver and spleen tissues of KO mice. In addition, the Results of IF assay suggested that GPR183 receptor induced the migration of spleen B lymphocytes from the central to the peripheral region of the follicles, and reduced the secretion of IL-17 and IL-22 by the spleen lymphocytes (P < 0.05). We also found that GPR183 KO mice significantly alleviated the process of acute liver injury in mice (P < 0.01). Conclusion A GPR183 knockout mouse model is successfully established, which provides a stable animal model for the further study of GPR183 gene function in vivo. Moreover, the knockout of GPR183 receptor can alleviate acute liver injury in mice.