Construction and Identification of Human IL-17RA-Deficient Cell Model

Objective: To construct and identify the human IL-17RA-deficient cell model.Methods: According to the published target sequence of IL-17RA,oligonucleotides were designed and synthesized,annealed,and inserted into pSilencer 2.1-U6 subsequently.The recombinant plasmid was confirmed by restriction enzyme digestion and DNA sequencing.The recombinant eukaryotic expression vector was transfected into 293T cells,IL-17RA mRNA and protein levels were detected by the real-time fluorescence quantitative PCR and flow cytometry respectively.Results: The construction of IL-17RA shRNA expression vector and IL-17RA-deficient cells were confirmed successfully by the restriction digestion and real-time quantitative PCR analysis respectively.Flow cytometry showed that IL-17RA expression in IL-17RA-deficient cells was significantly lower than in wild-type cells.Conclusion: The construction of IL-17RA-deficient cell model lays the foundation to study the function of IL-17 in osteoarthritis and to design new drug for osteoarthritis treatment.