| 摘要: |
| [摘要]目的：验证微小RNA410（miR-410）对白介素-13（IL-13）mRNA的靶向调控作用。方法：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-13 mRNA的表达。 |
| 关键词: 微小RNA410 白介素-13 双荧光素酶报告基因 |
| DOI：doi:10.13407/j.cnki.jpp.1672-108X.2021.04.001 |
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| 基金项目:国家自然科学基金,编号81701587。 |
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| Targeted Regulation Role of MicroRNA410 in Expression of Interleukin-13 Detected by Dual-Luciferase Reporter System |
| Jin Rong, Liu Dongyun, Hu Sujuan, Lin Rongjun |
| (The Affiliated Hospital of Qingdao University, Shandong Qingdao 266000, China) |
| Abstract: |
| [Abstract] 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 into 293T 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 dual-luciferase 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. |
| Key words: microRNA410 interleukin-13 dual-luciferase reporter system |
