〔Abstract〕 To construct RAW264.7 cell line with G-protein-coupled receptor 41(gpr41) gene knockout by CRISPR-Cas9 gene editing technique. Lentivirus was used to transfect Cas9 protein into macrophages. gRNA targetinggpr41 gene was designed to target the functional region ofgpr41 gene, and pLenticrisprV2 recombinant plasmid transformed DH5α competent cells were constructed to screen out recombinant sequencing to verify the effectiveness of gRNA. The plenticrisprV2-gpr41-gRNA recombinant plasmid was transfected with the macrophage line RAW264.7 using transfection reagent. The expression ofgpr41 gene was detected by Western blot. Sequencing results confirmed the successful construction of pLenticrisprv2-gpr41-gRNA recombinant plasmid.Western blot selection of recombinant plasmid pLenticrisprv2-gpr41-gRNA 3 transfected RAW264.7 cell No. 13 monoclonal strain showed no expression of GPR41 protein. CRISPR-Cas9 gene editing technique was used to successfully construct a macrophage line withgpr41 gene knockout,which laid the foundation for studying the mechanism ofgpr41 gene in RAW264.7 cells.