鸡miRNA-205b靶向调控CD24基因的双荧光素酶验证

Functional Verification of miRNA-205b Targeted CD24 Gene by Double Luciferase Reporter System

miRNA靶基因预测软件发现,热稳定抗原分子24(CD24)基因是miR-205b的候选靶基因,为了进一步研究miR-205b在鸡热应激中的作用,将CD24野生型、突变型(CD24-WT/MT)和miR-205b mimic及mimic NC共转染DF-1细胞后,检测双荧光素酶活性。结果显示:当共转染CD24-WT时,miR-205b mimic组显著的抑制荧光素酶的活性,而共转染CD24-MT时,mimic NC组和miR-205b mimic组差异不显著。表明miR-205b能够靶向CD24的3′UTR区域,抑制其表达,结果为深入分析miR-205b在鸡热应激中的功能和作用机制奠定基础。

The target gene prediction software was found that CD24 molecule(CD24)gene was a candidate target gene for miR-205b,to further study and explore the function of miR-205b in chicken heat stress,dual fluorescence was detected after co-transfection of CD24 wild-type and mutan-type(CD24-WT/MT)with miR-205b mimic and mimic NC in DF-1 cells.The results showed that the mimic group significantly inhibited the luciferase activity,while when CD24-MT was co-transfected,the differences between the mimic NC group and the miR-205b mimic group were not significant.The results demonstrated that miR-205b could regulate CD24 expression by target binding to its 3'UTR.This study could lay a good foundation for studying the function and mechanism of miR-205b in chicken heat stress.