Functional analysis of Smg6/Est1 in stem cell maintenance and differentiation

Smg6/Est1在干细胞维持和分化中的功能分析

• 批准号: 264350668
• 负责人: Professor Dr. Zhao-Qi Wang
• 金额: --
• 依托单位: Leibniz-Institut für Alternsforschung - Fritz-Lipmann-Institut e.V. (FLI)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/264350668?language=en
• 关键词: Functional; analysis; Smg6; Est1; stem

Est1, ever shorter telomere 1, is originally discovered as a co-factor of the telomerase machinery. Loss of Est1 in yeast causes telomere shortening and cell senescence. Vertebrate Est1, Smg6, is also involved in non-sense mediated mRNA decay (NMD) degrading aberrant mRNAs with premature termination codon (PTC), which may be produced by alternative splicing (AS). AS events have a profound role in various cellular processes, including cell proliferation and cell death as well as embryonic development. Over 90% of human multi-exonic genes are alternatively spliced; over 60% of human disease-causing mutations affect splicing rather than changing coding sequences. It is hypothesised that NMD balances the reservoir of gene isoforms, and contributes to the biological processes of development and tissue homeostasis. Our recent study shows that Smg6 deletion causes early embryonic lethality due to blocked differentiation of embryonic stem cells. How Smg6 regulates stem cell self-renewal and differentiation and development remains unknown. The objectives of the project: (1) To study the biological function of Smg6; (2) To characterize the Smg6-NMD specific targets in regulating stem cell maintenance and differentiation; (3) To understand the functions of Smg6 in stem cell maintenance and ageing using mouse and cellular models.This project will benefit the interesting preliminary results and the expertise of our lab in mouse model and stem cells. We will employ the following approaches: (1) The generation and characterization of mice, ES and somatic cells, with Smg6 knock-out alleles; (2) RNA-seq and PAR-CLIP sequencing to identify Smg6 specific targets; (3) in vitro and in vivo differentiation assays; (4) Deletion of Smg6 in specific stem cell compartment, i.e. central nervous system. This study will advance our understandings of the stem cell maintenance, tissue homeostasis, regeneration and healthy ageing.

Est 1，更短的端粒1，最初被发现作为端粒酶机制的辅助因子。酵母中Est 1的缺失导致端粒缩短和细胞衰老。脊椎动物EST 1（Smg 6）也参与了无义介导的mRNA衰变（NMD），降解可能由选择性剪接（AS）产生的具有提前终止密码子（PTC）的异常mRNA。AS事件在各种细胞过程中具有深远的作用，包括细胞增殖和细胞死亡以及胚胎发育。超过90%的人类多外显子基因是选择性剪接的;超过60%的人类致病突变影响剪接而不是改变编码序列。据推测，NMD平衡基因亚型的库，并有助于发育和组织稳态的生物学过程。我们最近的研究表明，Smg 6缺失导致早期胚胎死亡，由于胚胎干细胞的分化受阻。Smg 6如何调节干细胞的自我更新和分化与发育仍然未知。该项目的目标：（1）研究Smg 6的生物学功能;（2）研究Smg 6-NMD在干细胞维持和分化中的特异性靶点;（3）利用小鼠和细胞模型研究Smg 6在干细胞维持和衰老中的作用。我们将采用以下方法：（1）产生和表征具有Smg 6敲除等位基因的小鼠、ES和体细胞;（2）RNA-seq和PAR-CLIP测序以鉴定Smg 6特异性靶点;（3）体外和体内分化测定;（4）在特定干细胞区室（即中枢神经系统）中缺失Smg 6。这项研究将促进我们对干细胞维持、组织稳态、再生和健康衰老的理解。