〔Abstract〕 Objective To establish a Tohoku hospital pediatrics-1(THP-1) cell line with G protein-coupled receptor108(GPR108) deletion and explore its functions. Methods According to the requirements of the clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system, two single guide RNAs(sgRNA1 and sgRNA2) paring to the flanking fragments of human GPR108 gene were designed and synthesized. The two oligonucleotides were inserted in the pL-CRISPR.EFS.GFP vector to generate the new recombinant vectors(pL-CRISPR.EFS.GFP-sgRNA1 and pL-CRISPR.EFS.GFP-sgRNA2). The recombinant vectors and packaging plasmids(pMD2.G and psPAX2), were co-transfected into 293T cells to generate virus for infecting THP-1 cells. The GFP+cells were screened and isolated in 96-well culture plates by flow cytometry to obtain single-cell clones. PCR and Western blot were used to detect whether GPR108 was successfully knocked out in THP-1 cells. Both GPR108+/+and GPR108~(-/-) THP-1 cells were treated with lipopolysaccharide(LPS). Interleukin 8(IL-8) derived from the THP-1 cells, which were treated by LPS, was detected with Western blot and cytometric bead array(CBA) analysis. Results The recombinant lentiviral vector pL-CRISPR.EFS.GFP-sgRNA was successfully constructed and single-cell clone F9 was obtained by flow cytometric sorting after transfection of THP-1 cells. PCR and Western blot both confirmed that F9 was a GPR108~(-/-) THP-1 single-cell clone. LPS stimulated GPR108~(-/-) and GPR108+/+THP-1 cells, both Western blot and CBA results showed a significant decrease in IL-8 synthesis and secretion in GPR108~(-/-) THP-1 cells. Conclusion The GPR108~(-/-) THP-1 cell clone is successfully obtained based on the CRISPR/Cas9 system. GPR108 deletion in THP-1 cells treated by LPS leads to a decrease of IL-8 expression and secretion. It lays the foundation for further research on the molecular mechanisms of GPR108 in the immune inflammatory response.