中国实验动物学报2024，Vol.32Issue(4)：444-450,7.DOI:10.3969/j.issn.1005-4847.2024.04.004
巨噬细胞特异性敲除KLF2基因小鼠模型构建与鉴定
Construction of macrophage-specific KLF2 gene knockout mice
摘要
Abstract
Objective To establish a macrophage-specific KLF2 gene knockout mouse model,and explore the regulatory effect of KLF2 on the macrophage inflammatory response.Methods KLF2flox/+mice were constructed using CRISPR/Cas9 gene editing technology.Target genotype mice were obtained by breeding with Lyz2-Cre+/+mice and screening genotypes through polymerase chain reaction,and KLF2 knockout efficiency was verified using genotyping,qRT-PCR,and Western Blot.Bone marrow-derived macrophages(BMDMs)were isolated and cultivated and mRNA levels of inflammation-related factors in lipopolysaccharide-induced BMDMs were detected.Results A KLF2flox/flox/Lyz2-Cre+mouse model was established.KLF2 mRNA and protein levels in mouse bone marrow and BMDMs were significantly lower in KLF2 knockout mice compared with the findings in control mice,while expression levels of KLF2 in the heart,liver,and kidney showed no significant changes compared with levels in the control group.There were no significant differences in body weight,diet,drinking water,and appearance between the two groups.The group receiving lipopolysaccharide stimulation showed significantly reduced interleukin(IL)-6 mRNA expression and significantly increased IL-1,iNOS,and CD86 mRNA expression in KLF2-deficient BMDMs compared with the control group.Conclusions We constructed a macrophage-specific KLF2 knockout mouse model,thus laying the foundation for further research on the regulatory effect and mechanism of macrophage KLF2 in clinical inflammatory-related diseases.关键词
Kruppel样因子2/特异性敲除小鼠/Cre/LoxP系统/巨噬细胞/动脉粥样硬化Key words
KLF2/specific knockout mice/Cre/LoxP system/macrophages/atherosclerosis分类
生物科学引用本文复制引用
孟雪,王新洲,高水波,吴鸿..巨噬细胞特异性敲除KLF2基因小鼠模型构建与鉴定[J].中国实验动物学报,2024,32(4):444-450,7.基金项目
国家自然科学基金(81473453),河南省科技研发计划联合基金(222301420088),河南省高校重点科研项目(23A360013). Funded by the National Natural Science Foundation of China(81473453),Henan Provincial Science and Technology Research and Development Plan Joint Fundation(222301420088),Key Scientific Research Projects of Universities in Henan Province(23A360013). （81473453）
