Objective To establish a macrophage-specific KLF2 gene knockout mouse model, and explore the regulatory effect of KLF2 on macrophage inflammatory response. Methods The KLF2flox/+mice were constructed using CRISPR/Cas9 gene editing technology. By breeding with Lyz2-Cre+/+ mice and screening genotypes through PCR, the target genotype mice were obtained, and the KLF2 knockout efficiency was verified using genotyping, qPCR and western blotting. BMDMs were Separated and cultivated, and the mRNA levels of inflammation-related factors in LPS-induced BMDMs were detected. Results A KLF2flox/flox/ Lyz2-Cre+ mouse model was established. The levels of KLF2 mRNA and protein in mouse bone marrow and BMDMs were significantly lower than those in the control group, while the expression of KLF2 in heart, liver, and kidney showed no significant changes compared to the control group. No significant differences in body weight, diet, drinking water and appearance were found between the two groups. Under the stimulation of LPS, the expression level of IL-6 mRNA in KLF2 deficient BMDMs was significantly lower than those in the control group, IL-1, iNOS, and CD86 mRNA were significantly higher than those in the control group. Conclusions The macrophage-specific knockout KLF2 mouse model was constructed, laying the foundation for further research on the regulatory effect and mechanism of macrophage KLF2 on clinical inflammatory-related diseases.