摘要
目的:验证微小RNA410(miR-410)对白细胞介素-13(IL-13) mRNA的靶向调控作用。方法:采用生物信息学软件预测miR-410与IL-13 mRNA 3’端非翻译区存在结合位点,设计并合成包含miR-410结合序列及突变序列的IL-13基因序列片段,分别构建野生型和突变型双荧光素酶基因载体,与miR-410和阴性对照序列共同转染293T细胞,检测转染后荧光素酶的活性,验证miR-410对IL-13 mRNA表达的影响。结果:生物信息学显示,IL-13 mRNA存在与miR-410的结合位点。基因测序结果显示,野生型和突变型荧光素酶报告基因载体构建成功。通过双荧光素酶报告系统发现,miR-410能够结合野生型IL-13 mRNA荧光素酶基因序列从而抑制其表达,而对突变型IL-13 mRNA的荧光素酶表达无影响(P<0.05)。结论:miR-410可以靶向调控IL-13mRNA的表达。
Objective: To verify the targeted regulation role of microRNA410( miR-410) on expression of interleukin-13( IL-13 mRNA. Methods: The binding sites of miR-410 in 3’untranslated region( 3’UTR) of IL-13 mRNA were predicted by the bioinformatics software. IL-13 gene fragments binding miR-410 and mutant sequences were designed and synthesized. The wild and mutant luciferase reporter gene carriers were respectively constructed. miR-410 mimics or negative control of mimics were respectively co-transfected into293 T cells with wild or mutant luciferase reporter gene. The luciferase activity was detected to observe the effects of miR-410 on expression of IL-13 mRNA. Results: Bioinformatics analysis showed that there was binding site located in 3’UTR of IL-13 mRNA. The electrophoresis showed that the wild and mutant luciferase reporter gene carriers were constructed successfully. Through the dualluciferase reportier system,it was found that miR-410 could bind the wild-type IL-13 mRNA luciferase gene sequence and inhibit its expression,but had no effect on the mutant IL-13 mRNA luciferase expression( P<0. 05). Conclusion: IL-13 mRNA can be targeted by miR-410.
作者
金蓉
刘冬云
胡素娟
林荣军
Jin Rong;Liu Dongyun;Hu Sujuan;Lin Rongjun(The Affiliated Hospital of Qingdao University,Shandong Qingdao 266000,China)
出处
《儿科药学杂志》
CAS
2021年第4期1-4,共4页
Journal of Pediatric Pharmacy
基金
国家自然科学基金,编号81701587。
