C-C趋化因子受体6基因敲除小鼠的构建及鉴定OA

Objective To establish a C-C motif chemokine receptor 6(CCR6)homozygous knockout mouse model in order to provide a crucial animal model foundation for subsequent in vivo functional studies.Methods Ccr6-/-mice were generated using CRISPR-Cas9 technology.Genomic DNA was extracted from mouse tails,with genotyp-ing performed by PCR and agarose gel electrophoresis.Pathological morphology of major organs(heart,liver,lung,kidney)was assessed through HE staining.Western blot was used to analyze CCR6 protein expression in blood,spleen,and bone marrow.To analyze the impact of CCR6 gene knockout on the proportion of major immune cell populations,the ratio of T cells and macrophages in the mouse spleen was detected using flow cytometry.Re-sults The results of agarose gel electrophoresis demonstrated that mice exhibiting a single specific band at the 307 bp position upon primer-based identification were confirmed as Ccr6-/-mice.HE staining revealed no significant histopathological differences between Ccr6+/+and Ccr6-/-mice.Western blot demonstrated near-complete absence of CCR6 protein in target tissues.Flow cytometry results demonstrated that CCR6 gene deletion significantly in-creased the proportion of CD8⁺T cells,while the ratios of both CD4⁺T cells and macrophages remained unaltered.Conclusion A Ccr6-/-mouse model is established using CRISPR-Cas9 technology,serving as an essential tool for elucidating CCR6's regulatory role in tumor proliferation.