期刊
STEM CELL RESEARCH & THERAPY
卷 10, 期 -, 页码 -出版社
BMC
DOI: 10.1186/s13287-019-1270-5
关键词
lncRNA; Snhg3; mESCs; Self-renewal; Pluripotency
资金
- National Key R&D Program of China [2017YFA0102801, 2017YFC1001901]
- National Nature Science Foundation of China [31671540, 31601179]
- Natural Science Foundation of Guangdong Province [2015A020212005, 2016A030310209]
- Guangzhou Science and Technology Project [201803010020]
- Fundamental Research Funds of State Key Laboratory of Ophthalmology [30306020240020307]
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.
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