Identification of Cis-element and Trans-factors Functioning
发布时间：2022-11-05 14:37
研究背景越来越多的下一代测序数据表明人类基因组中绝大多数区域都被转录。在这些转录产物中,lncRNA是最丰富的一类RNA,它们通过与RNA,DNA和蛋白质相互作用发挥着不同的作用和多种功能。与mRNA类似,lncRNA由RNA po1Ⅱ转录,在5’端有帽子结构,其中大多数具有剪接事件和polyA尾。根据定义,lncRNAs不编码蛋白质,这是lncRNAs和mRNAs之间的根本区别。对于在细胞中的定位,所有mRNA都被转运到细胞质中进行翻译,而lncRNA可能定位在细胞核、细胞质或两个区域中。经过剪接的mRNA向细胞质的有效转运输出显然是由于在剪接过程中将TREX复合物募集到mRNA的5’末端,相反,尽管同样有剪接事件,许多经过剪接的lncRNA仍滞留在细胞核中。一些研究已经提供了关于lncRNAs核滞留的一些线索。例如,lncRNA FIRRE含有156bp重复RNA结构域,这些结构域对于FIRRE定位于靶染色体上至关重要。类似地,XIST 5’重复元件有助于XIST RNA在X染色体上定位。在这两种情况下,敲低hnRNPU导致这些RNA从染色体上脱离。另一项研究表明,Malat1含有...
【文章页数】：101 页
【学位级别】：博士
【文章目录】：
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
本文编号：3702787
【文章页数】：101 页
【学位级别】：博士
【文章目录】：
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
本文编号：3702787
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