Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/34724
Title: Roles of Long Non-Coding RNAs in Human Embryonic Stem Cell Pluripotency and Neural Differentiation
Authors: NG SHI YAN
Keywords: Pluripotency, Neurogenesis, lncRNAs, Long non-coding RNAs.
Issue Date: 23-Mar-2012
Source: NG SHI YAN (2012-03-23). Roles of Long Non-Coding RNAs in Human Embryonic Stem Cell Pluripotency and Neural Differentiation. ScholarBank@NUS Repository.
Abstract: Long non-coding RNAs (lncRNAs) are a recently discovered class of transcripts encoded within the human genome. LncRNAs have been proposed to be key regulators of biological processes, including stem cell pluripotency and neurogenesis. However, at present very little functional characterization of lncRNAs involved in differentiation has been carried out in human cells. In this thesis, functional characterization of lncRNAs in human development is addressed using human embryonic stem cells (hESCs) as a paradigm for pluripotency and neuronal differentiation. Human ESCs were robustly and efficiently differentiated into neurons, and expression of lncRNAs was profiled using a custom-designed microarray. Some hESC-specific lncRNAs involved in pluripotency maintenance were identified, and shown to physically interact with SOX2, and PRC2 complex component, SUZ12. Using a similar approach, we identified lncRNAs required for neurogenesis. Knockdown studies indicated that loss of any of these lncRNAs blocked neurogenesis, and immunoprecipitation studies revealed physical association with REST and SUZ12. In particular, a neuronal lncRNA, RMST, was found to be essential for neurogenesis. Knockdown of RMST in human neural stem cells prevented neurogenesis. RNA pulldown and RNA immunoprecipitation indicated that RMST physically associated with the RNA-binding protein hnRNPA2B1 and the transcription factor SOX2. Perturbation studies, followed by genome-wide transcriptional profiling indicated that RMST and SOX2 co-regulate a large pool of targets. Interestingly, knockdown of RMST resulted in reduced SOX2 occupancy at its target gene promoters, suggesting that RMST may alter SOX2 binding to chromatin during neurogenesis. Together, this study represents important evidence for an indispensable role of lncRNAs in human brain development.
URI: http://scholarbank.nus.edu.sg/handle/10635/34724
Appears in Collections:Ph.D Theses (Open)

Show full item record
Files in This Item:
File Description SizeFormatAccess SettingsVersion 
NgSY_Part1.pdf12.3 MBAdobe PDF

OPEN

NoneView/Download
NgSY_Part1B.pdf81 kBAdobe PDF

OPEN

NoneView/Download
NgSY_Part2.pdf6.65 MBAdobe PDF

OPEN

NoneView/Download
NgSY_Part3.pdf5.76 MBAdobe PDF

OPEN

NoneView/Download
NgSY_Part4.pdf12.77 MBAdobe PDF

OPEN

NoneView/Download

Page view(s)

305
checked on Dec 2, 2017

Download(s)

1,125
checked on Dec 2, 2017

Google ScholarTM

Check


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.