The long non-coding RNA Snhg3 is essential for mouse embryonic stem cell self-renewal and pluripotency
The long non-coding RNA Snhg3 is essential for mouse embryonic stem cell self-renewal and pluripotency
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长链非编码RNA Snhg3对于小鼠胚胎干细胞的自我更新和多能性至关重要
DOI：
10.1186/s13287-019-1270-5
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发表时间：
2019-05-31
影响因子：
7.5
通讯作者：
Huang, Junjiu
中科院分区：
文献类型：
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作者：
Lu, Weisi;Yu, Jianping;Huang, Junjiu
背景小核仁RNA宿主基因3（Snhg3）是一种长非编码RNA（lncRNA），已被证明参与某些癌症的肿瘤发生。然而，人们对其在胚胎干细胞 (ESC) 中的作用知之甚少。方法在这里，我们通过功能丧失（敲低）和功能获得（过表达）方法研究了 Snhg3 在小鼠 ESC (mESC) 中的作用。使用碱性磷酸酶染色、二次集落形成、碘化丙啶染色、蛋白质印迹和定量逆转录聚合酶链反应（qRT-PCR）来评估自我更新能力，而进行免疫荧光、qRT-PCR和拟胚体形成来检查多能性。此外，根据形态变化、囊胚率和改变的多能性标记（Nanog、Oct4）表达来确定 Snhg3 对小鼠胚胎发育的影响。此外，通过染色质免疫沉淀 qPCR、qRT-PCR、亚细胞分级分离和 RNA 免疫沉淀评估了 Snhg3 与关键多能性因子之间的关系。最后，应用RNA pull-down和质谱分析来探索mESCs中Snhg3的潜在相互作用蛋白。结果我们证明Snhg3对于mESCs的自我更新和多能性维持至关重要。此外，Snhg3 敲低会破坏小鼠早期胚胎发育。从机制上讲，Snhg3与Nanog和Oct4形成正反馈网络，并在mESC中鉴定出126个Snhg3相互作用蛋白。结论Snhg3对于mESC的自我更新和多能性以及小鼠早期胚胎发育至关重要。
BackgroundSmall nucleolar RNA host gene 3 (Snhg3) is a long non-coding RNA (lncRNA) that was shown to participate in the tumorigenesis of certain cancers. However, little is known about its role in embryonic stem cells (ESCs).MethodsHere, we investigated the role of Snhg3 in mouse ESCs (mESCs) through both loss-of-function (knockdown) and gain-of-function (overexpression) approaches. Alkaline phosphatase staining, secondary colony formation, propidium iodide staining, western blotting, and quantitative reverse transcription polymerase chain reaction (qRT-PCR) were used to access self-renewal capacity, whereas immunofluorescence, qRT-PCR, and embryoid body formation were performed to examine pluripotency. In addition, the effect of Snhg3 on mouse embryonic development was determined based on the morphological changes, blastocyst rate, and altered pluripotency marker (Nanog, Oct4) expression. Moreover, the relationship between Snhg3 and key pluripotency factors was evaluated by chromatin immunoprecipitation qPCR, qRT-PCR, subcellular fractionation, and RNA immunoprecipitation. Finally, RNA pull-down and mass spectrometry were applied to explore the potential interacting proteins of Snhg3 in mESCs.ResultsWe demonstrated that Snhg3 is essential for self-renewal and pluripotency maintenance in mESCs. In addition, Snhg3 knockdown disrupted mouse early embryo development. Mechanistically, Snhg3 formed a positive feedback network with Nanog and Oct4, and 126 Snhg3-interacting proteins were identified in mESCs.ConclusionsSnhg3 is essential for mESC self-renewal and pluripotency, as well as mouse early embryo development.