請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/49483
標題: | 探索Rtl1於小鼠腦中的印記狀態及其角色 Investigating the Imprinting Status and Role of Rtl1 in the Mouse Brain |
作者: | Meng-Ying Lin 林孟穎 |
指導教授: | 黃憲松(Hsien-Sung Huang) |
關鍵字: | 表觀遺傳學,基因組印記,Dlk1-Dio3基因座,差異性甲基化區域,Rtl1,腦區特異印記, Epigenetics,Genomic imprinting,Dlk1-Dio3 locus,Differentially methylated region,Rtl1,Brain region-specific imprinting, |
出版年 : | 2016 |
學位: | 碩士 |
摘要: | 基因組印記是一個表觀遺傳學的現象:哺乳類中多數基因乃是由雙親的等位基因均等表達,但有些基因自父母雙方的等位基因表達的程度不相等,主要從父方表達或是主要從母方表達,這類基因稱為印記基因。印記基因在發育及細胞分化上扮演重要角色,因此,其失調可能會導致發育異常。Rtl1是一個編碼的印記基因,其所在的Dlk1-Dio3基因座位於小鼠染色體12號及人類染色體14號上。根據過往研究,Rtl1的過度表現為導致人類疾病「鏡─緒方症候群」症狀的可能原因之一,此疾病的病患具有鐘形胸腔、腹壁缺損、發育遲緩等症狀,亦有可能出現智力障礙。了解Rtl1的調控機制及其功能對此疾病的治療或許有所幫助。根據過往研究,在小鼠胚胎及胎盤中Rtl1為父方表達,但在我們及其他研究團隊的RNA定序資料中發現,在部分腦區Rtl1為母方表達。RTL1目前已知其在胎盤中的功能是維持胎兒微血管,但在腦中的功能仍鮮為人知,我們的研究偏重Rtl1腦中所扮演的角色,及其功能是否與特殊印記狀態有關。藉由DNA定序,我們發現Rtl1在不同腦區中其印記狀態有所差別,而此差異可能與Rtl1在不同腦區的表現量多寡有關。我們嘗試找出調控Rtl1等位基因表達的機制,為此測驗了在此基因座的三個差異性甲基化區域的甲基化程度。此外,為了於活體內研究RTL1的功能,我們利用CRISPR技術製造了Rtl1基因剃除小鼠。此研究可幫助人們了解更多關於Rtl1的調控機制及其在腦中所扮演的角色。 Genomic imprinting is an epigenetic phenomenon that some genes called imprinted genes are expressed in a parent-of-origin-specific manner. Imprinted genes play an important role during development and differentiation. Therefore, dysfunction of imprinted genes may cause developmental problem. Retrotransposon-like 1 (Rtl1) is a protein-coding imprinted gene, and it is located in the Dlk1-Gtl2 locus on mouse chromosome 12 and on human chromosome 14. Previous studies have indicated that overexpression of Rtl1 might cause the symptoms of a human disorder named Kagami-Ogata syndrome. Patients of this disorder have symptoms such as coat-hanger appearance of the ribs, abdominal wall defects, developmental delay and/or intellectual disability. Understanding the mechanism of Rtl1 modulation and the role it plays may be helpful to treat this syndrome. Rtl1 is known to be paternally expressed in mice fetus and placenta. However, not only our group’s but also other groups’ RNA-Seq data show that Rtl1 is maternally expressed in some brain regions. RTL1 maintains the fetal capillaries in the feto-maternal interface of placenta, but the role of RTL1 in the brain is totally unknown. We are studying the function of RTL1 and try to figure out whether its function is related to its imprinting status in the brain. Through Sanger sequencing, we confirmed that imprinting statuses of Rtl1 are distinct in different brain regions, and this phenomenon may be associated with Rtl1 expression level. To find out the mechanisms which regulate the Rtl1 imprinting status in distinct brain regions, we also determined the methylation level in three differentially methylated regions of this locus. To understand the physiological role of RTL1 in the brain in vivo, we generated the Rtl1 knockout mice through CRISPR/Cas9 system. This study can help us knowing the mechanism which modulates Rtl1 and the role of RTL1 in the brain in more details. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/49483 |
DOI: | 10.6342/NTU201603098 |
全文授權: | 有償授權 |
顯示於系所單位: | 腦與心智科學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-105-1.pdf 目前未授權公開取用 | 9.95 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。