研究背景越來越多的下一代測序數(shù)據(jù)表明人類基因組中絕大多數(shù)區(qū)域都被轉(zhuǎn)錄。在這些轉(zhuǎn)錄產(chǎn)物中,lncRNA是最豐富的一類RNA,它們通過與RNA,DNA和蛋白質(zhì)相互作用發(fā)揮著不同的作用和多種功能。與mRNA類似,lncRNA由RNA po1Ⅱ轉(zhuǎn)錄,在5’端有帽子結(jié)構(gòu),其中大多數(shù)具有剪接事件和polyA尾。根據(jù)定義,lncRNAs不編碼蛋白質(zhì),這是lncRNAs和mRNAs之間的根本區(qū)別。對于在細(xì)胞中的定位,所有mRNA都被轉(zhuǎn)運到細(xì)胞質(zhì)中進(jìn)行翻譯,而lncRNA可能定位在細(xì)胞核、細(xì)胞質(zhì)或兩個區(qū)域中。經(jīng)過剪接的mRNA向細(xì)胞質(zhì)的有效轉(zhuǎn)運輸出顯然是由于在剪接過程中將TREX復(fù)合物募集到mRNA的5’末端,相反,盡管同樣有剪接事件,許多經(jīng)過剪接的lncRNA仍滯留在細(xì)胞核中。一些研究已經(jīng)提供了關(guān)于lncRNAs核滯留的一些線索。例如,lncRNA FIRRE含有156bp重復(fù)RNA結(jié)構(gòu)域,這些結(jié)構(gòu)域?qū)τ贔IRRE定位于靶染色體上至關(guān)重要。類似地,XIST 5’重復(fù)元件有助于XIST RNA在X染色體上定位。在這兩種情況下,敲低hnRNPU導(dǎo)致這些RNA從染色體上脫離。另一項研究表明,Malat1含有... 

【文章頁數(shù)】：101 頁

【學(xué)位級別】：博士

【文章目錄】：
ABSTRACT(English)
ABSTRACT(摘要)
INTRODUCTION
    Long noncoding RNA (LncRNA) Introduction
    Mechanism of RNA localization
    Localization of LncRNAs
    Importance of lncRNA localization
    Techniques Used for LncRNA Localization
    Nuclear Retention of lncRNAs
MATERIALS AND METHODS
    Constructs and Antibodies
    Cell Culture and Transfection
    Fluorescence in situ Hybridization and Immunofluorescence
    Mini Nuclear Extract Preparation
    In Vitro RNP Purification
    Mass Spectrometry Analysis
    RNA Sequencing Analysis(RNA-seq)
RESULTS
    Nuclear Localization of MEG3 Reporter Transcript
    Identification of nuclear retention element in MEG3
    Nuclear Retention of Cytoplasmic RNAs by MEG3 NRE
    NRE Associated LncRNPs Purification
    Knockdown of U1snRNP Led to accumulation of MEG3 reporter transcripts in cytoplasm
    Knockdown of U1snRNP Led to cytoplasmic accumulation of LncRNAs
DISCUSSION
REFERENCES
LITERATURE REVIEW
    Introduction
    Identification and Characterization of LncRNAs
    LncRNA Databases
    LncRNA Regulatory Roles
    LncRNA Role in Cancer Stem Cells CSCs
    BCSCs Regulation by LncRNAs
    BCSCs Therapeutic LncRNAs
    REFERENCES
APPENDIX
ACKNOWLEDGEMENTS



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