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        體細胞重編程中miRNA的鑒定及功能研究

        2016-05-30 18:24:04程紅
        科技資訊 2016年21期

        程紅

        摘 要:該研究針對非編碼RNA研究中的核心科學(xué)問題,研究體細胞重編程微RNA功能及調(diào)控網(wǎng)絡(luò)。研究的5年預(yù)期目標是:利用分子生物學(xué)、細胞生物學(xué)、生物化學(xué)和信息生物學(xué)手段,完成重編程和體細胞重編程和多能性細胞分化多個階段的微RNA的表達譜,鑒定出在以上多個階段表達變化顯著的微RNA。找到影響重編程或多能干細胞分化過程的幾種微RNA及鑒定其靶基因,并揭示這些基因在上述過程的作用。鑒定到在轉(zhuǎn)錄后加工水平和核質(zhì)轉(zhuǎn)運過程受調(diào)控的微RNA,闡明這些特定微RNA的作用機理,揭示其核質(zhì)轉(zhuǎn)運變化與重編程和多能性細胞分化之間的關(guān)聯(lián),闡明其分子機制。闡明上述相關(guān)微RNA在胚胎發(fā)育、細胞分化和重編程過程中的作用及其自身生物合成規(guī)律,并揭示其在器官發(fā)育中的作用。近3年,已建立體細胞重編程和多能性細胞分化及體細胞轉(zhuǎn)分化的研究系統(tǒng)。采用核移植技術(shù),建立了可用于研究體細胞重編程中miRNA功能的新系統(tǒng)——來自孤雄囊胚的單倍體胚胎干細胞系。已經(jīng)建立了研究miRNA核質(zhì)轉(zhuǎn)運和轉(zhuǎn)錄后加工調(diào)控的研究體系。制作了miRNA敲除小鼠。系統(tǒng)鑒定了體細胞重編程過程中差異表達的miRNA。系統(tǒng)鑒定了體細胞向肝細胞轉(zhuǎn)分化過程中顯著差異表達的miRNA。發(fā)現(xiàn)了RNA結(jié)合蛋白在轉(zhuǎn)錄后加工水平調(diào)控的特定miRNA的表達。發(fā)現(xiàn)在哺乳動物細胞核內(nèi)存在一種讀碼機制,可能參與調(diào)控非編碼RNA的核質(zhì)轉(zhuǎn)運。揭示了miR-126在小鼠胚胎血管新生和血管完整性維持中的作用,探究了其發(fā)揮作用的分子機制。

        關(guān)鍵詞:非編碼RNA 細胞重編程 干細胞分化 微RNA 核質(zhì)轉(zhuǎn)運

        Abstract: This proposal is focused on microRNAs that are involved in cell reprogramming. Using molecular biological, cell biological and biochemical methods, we plan to establish the expression profile of microRNAs at multiple stages of cell reprogramming and stem cell differentiation, and identify microRNAs whose expression level change significantly during these processes. Through these studies, we hope to identify microRNAs that play important regulatory roles in these processes and identify their target genes. We also plan to identify microRNAs whose expression levels are regulated at post-transcriptional levels and investigate the molecular mechanism. In the recent three years, we have established the systems that are suitable for study the processed of cell reprogramming and differentiation. Using nuclear transfer technique, we have established the systems for generation of mice by oocyte injection of androgenetic haploid embryonic stem cells that can be used for function of microRNA. We have also set up the research system for study of nucleocytoplasmic transportation and processing of microRNAs. We have obtained a line of microRNA knockout mouse. According to our plan, we have identified multiple microRNAs whose expression level changed significantly during cell reprogramming and trans-differentiation of somatic cells to hepatocellular cells. We have investigated the function and mechanism for mir-126 in regulating vescular integrity. In the past three years, we have published 10 research papers including one paper that was published in Cell.

        Key Words: Non-coding RNA; Cell reprogramming; Stem cell differentiation; MicroRNA; Nucleocytoplasmic transportation

        閱讀全文鏈接(需實名注冊):http://www.nstrs.cn/xiangxiBG.aspx?id=49703&flag=1

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