安徽医科大学学报2026，Vol.61Issue(3)：409-415,7.DOI:10.19405/j.cnki.issn1000-1492.2026.03.004
C-C趋化因子受体6基因敲除小鼠的构建及鉴定
Construction and identification of C-C motif chemokine receptor type 6 gene knockout mice
摘要
Abstract
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.关键词
C-C趋化因子受体6/基因敲除/CRISPR-Cas9/基因型鉴定/聚合酶链式反应/蛋白表达Key words
C-C motif chemokine receptor 6/gene knockout/CRISPR-Cas9/genotyping/polymerase chain reac-tion/protein expression分类
医药卫生引用本文复制引用
吴瑶瑶,张蕊,魏伟..C-C趋化因子受体6基因敲除小鼠的构建及鉴定[J].安徽医科大学学报,2026,61(3):409-415,7.基金项目
国家自然科学基金项目(编号:82403663) （编号:82403663）
安徽省高校自然科学研究重点项目(编号:2024AH050776) （编号:2024AH050776）
安徽医科大学校科研基金项目(编号:2023xkj013) National Natural Science Foundation of China(No.82403663) （编号:2023xkj013）
Natural Science Research Proj-ect of Anhui Educational Committee(No.2024AH050776) （No.2024AH050776）
Scientific Research Project of Anhui Medical Univer-sity(No.2023xkj013) （No.2023xkj013）
