LncRNA Riken-201 and Riken-203 modulates neural development by regulating the Sox6 through sequestering miRNAs
LncRNA Riken-201 and Riken-203 modulates neural development by regulating the Sox6 through sequestering miRNAs
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LncRNA Riken-201 和 Riken-203 通过隔离 miRNA 调节 Sox6 来调节神经发育
DOI：
10.1111/cpr.12573
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发表时间：
2019-05-01
影响因子：
8.5
通讯作者：
Jiang，Hong
中科院分区：
文献类型：
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作者：
Zhang，Lei;Xue，Zhenyu;Jiang，Hong
目的长链非编码RNA（LncRNA）在发育过程中的表观遗传调控中发挥重要作用。在本研究中，我们发现通过选择性剪接，LncRNA C130071 C 03 Riken变体Riken-201（Riken-201）和Riken-203（Riken-203）都在大脑中高表达，并在神经分化过程中逐渐增加。材料和方法胚胎干细胞（ESCs）; RNA测序; mRNA、LncRNA和miRNA的基因表达;基因的过表达和RNA干扰;流式细胞术;真实的实时定量PCR;和Western blot。采用RNA pull-down试验和PCR检测与Rik-201和Rik-203连接的任何miRNA。荧光素酶法检测miRNA与Sox 6 mRNA的结合情况。结果Rik-201和Rik-203的表达抑制小鼠胚胎干细胞向神经细胞的分化。此外，Rik-201和Rik-203作为竞争性内源性RNA（ceRNA）分别抑制miR-96和miR-467 a-3 p的功能，并调节Sox 6的表达以进一步调节神经分化。Rik-203和Rik-201的敲除诱导高比率的脑发育迟缓。结论Rik-201和Rik-203在促进神经分化和脑发育中的功能作用，并阐明miRNA-Sox 6-相关的分子机制。
ObjectivesLong non‐coding RNAs (LncRNAs) play important roles in epigenetic regulatory function during the development processes. In this study, we found that through alternative splicing, LncRNA C130071C03Riken variants Riken‐201 (Riken‐201) and Riken‐203 (Riken‐203) are both expressed highly in brain, and increase gradually during neural differentiation. However, the function of Rik‐201 and Rik‐203 is unknown.Materials and methodsEmbryonic stem cells (ESCs); RNA sequencing; gene expression of mRNAs, LncRNAs and miRNAs; over‐expression and RNA interference of genes; flow cytometry; real‐time quantity PCR; and Western blot were used in the studies. RNA pull‐down assay and PCR were employed to detect any miRNA that attached to Rik‐201 and Rik‐203. The binding of miRNA with mRNA of Sox6 was presented by the luciferase assay.ResultsRepression of Rik‐201 and Rik‐203 inhibited neural differentiation from mouse embryonic stem cells. Moreover, Rik‐201 and Rik‐203 functioned as the competing endogenous RNA (ceRNA) to repress the function of miR‐96 and miR‐467a‐3p, respectively, and modulate the expression of Sox6 to further regulate neural differentiation. Knockout of the Rik‐203 and Rik‐201 induced high ratio of brain developmental retardation. Further we found that C/EBPβ might potentially activated the transcription of Rik‐201 and Rik‐203.ConclusionsThese findings identify the functional role of Rik‐201 and Rik‐203 in facilitating neural differentiation and further brain development, and elucidate the underlying miRNAs‐Sox6‐associated molecular mechanisms.