靶向鸡糖皮质激素受体基因的microRNA预测与鉴定OA北大核心CSTPCD

[Objectives]This study aimed to predict and screen microRNA that targeted the expression of chicken glucocorticoid receptor(GR).[Methods]The microRNA targeting chicken GR 3′UTR was predicted by TargetScan,PicTar and miRDB software,respectively,and the overlapping microRNA was selected as candidates for further experiment.The recombinant plasmid and mutant plasmid containing chicken GR 3′UTR were constructed,and microRNA binding to chicken GR 3′UTR was verified by double luciferase assay.The target genes of candidate microRNA were predicted by TargetScan and miRDB software,respectively,and the GO function of the predicted common target genes was analyzed by DAVID software.Western blot was used to detect the expression of GR protein after overexpression of candidate microRNA in chicken DF1 cells.[Results]miR124-3p,miR142-3p,miR204/211,miR183,miR18-5p,and miR181a/181b were the intersection microRNA of 3 softwares prediction targeting GR 3′UTR.Dual luciferase assay showed that miR142-3p,miR204/211 and miR18-5p targeted GR 3′UTR.The two kinds of software predicted that the target genes of three candidate microRNA contained GR and the enriched molecular functions were different.After overexpression of the three candidate microRNA in DF1 cell,miR142-3p and miR204/211 significantly decreased the expression of GR protein level(P<0.05).[Conclusions]miR142-3p,miR204/211 and miR18-5p target chicken GR 3′UTR,and miR142-3p and miR204/21 can reduce the expression level of GR in chicken DF1 cell.