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dx5在胚胎干细胞及重编程中的功能研究

he function of Ddx5 in embryonic stem cells and reprogramming

中文摘要英文摘要

目的:研究Ddx5在胚胎干细胞(Embryonic stem cell, ES cell)、诱导多能干细胞(induced pluripotent stem cell, iPS cell)中的作用以及在干细胞、重编程中对下游基因的选择性剪接作用。方法: PCR检测Ddx5在ES细胞及其分化过程中的表达水平,重编程过程中的基因表达变化;通过在ES细胞中过表达和敲低Ddx5,检测其对ES多能性的影响;在体细胞重编程中检测Ddx5的过表达和敲低引起的效率变化,利用流式细胞术和碱性磷酸酶染色检测重编程效率;最后利用PCR检测,分析Ddx5对下游基因的选择性剪接方式的影响。结果:Ddx5在ES细胞中富集并随分化降低。在ES细胞中过表达Ddx5后,Nanog、Eras等多能性基因表达上调,敲低则相反。Ddx5在重编程过程中表达上调,其过表达促进重编程,敲低则明显抑制。Ddx5过表达引起CD44、Hras等基因的选择性剪切变化。结论:Ddx5在ES细胞和重编程过程中都发挥着作用,Ddx5过表达引起选择性剪切变化。

Objective: To investigate the roles of Ddx5 in ES cell (Embryonic stem cell) and iPS cell (induced pluripotent stem cell) and to study the function of them in alternative splicing in ES cells and reprogramming process. Methods:Use PCR we examined the mRNA level of Ddx5 in ES cells and differentiation, and reprogramming. By overexpression or knockdown, we explored the role of Ddx5 in ES pluripotency maintenance. In addition, we examined the expression level of Ddx5 in reprogramming and investigated the effect of Ddx5 on reprogramming efficiency. Finally, we test the splicing changes of some target genes under Ddx5 overexpression using splicing PCR. Results: Ddx5 is enrich in ES cells and reduces during differentiation. The overexpression of Ddx5 in ES cells results in the up-regulation of pluripotency genes, and the knockdown of Ddx5 is opposite. Moreover, the expression of Ddx5 in reprogramming is upregulation and promotes efficiency of reprogramming. In addition, Ddx5 leads to the alternative splicing changes of CD44、Hras and so on. Conclusion:Ddx5 plays significant roles in ES cells and reprogramming. And overexpression of Ddx5 causes the alternative splicing changes.

郭菲菲、康岚

细胞生物学遗传学分子生物学

分子生物学ES重编程dx5dx17选择性剪接

Molecular biologyESreprogrammingDdx5Ddx17selective splicing

郭菲菲,康岚.dx5在胚胎干细胞及重编程中的功能研究[EB/OL].(2017-04-28)[2025-08-18].http://www.paper.edu.cn/releasepaper/content/201704-817.点此复制

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