Edinburgh Research Archive logo

Edinburgh Research Archive

University of Edinburgh homecrest
View Item 
  •   ERA Home
  • Edinburgh Medical School
  • Edinburgh Medical School thesis and dissertation collection
  • View Item
  •   ERA Home
  • Edinburgh Medical School
  • Edinburgh Medical School thesis and dissertation collection
  • View Item
  • Login
JavaScript is disabled for your browser. Some features of this site may not work without it.

Dissecting the function of MYEF2 in neural stem cells and glioblastoma

View/Open
McCarten2020.pdf (33.89Mb)
Date
27/06/2020
Item status
Restricted Access
Embargo end date
27/06/2021
Author
McCarten, Katrina Anne
Metadata
Show full item record
Abstract
Glioblastoma (GBM) is the most common malignant primary brain tumour in adults, with a median survival time of approximately ~15 months after diagnosis. GBM stem cells (GSCs) underpin GBM’s aggressiveness, resistance to radiation and chemotherapy, and disease recurrence. Glioblastoma stem cells (GSCs) are capable of tumour formation upon orthotopic transplantation into immunocompromised mice and share many properties of neural stem cells (NSCs). One feature common to both GSCs and NSCs is high expression of the transcription factor SOX2. SOX2 is known to be essential for GCS self-renewal. To understand further how SOX2 exerts its function in GSCs, the Pollard laboratory studied the interactome of SOX2. Using SICAP-MS – a method that looks at the on-chromatin interactors of the protein of interest – the Pollard laboratory have identified MYEF2 as a candidate interacting partner. MYEF2 contains multiple RNA binding domains and is expressed primarily in the brain and testis. Here I explored the function of MYEF2 within NSCs and GSCs. I confirm Myef2 is expressed within the brain, however was not specifically enriched in stem cell containing regions. CRISPR/Cas9 mediated knock-in of mCherry and HA to Myef2 was successfully carried out in mouse NSCs. We observed that MYEF2 is a nuclear protein which retains nuclear localisation and expression in both proliferative and quiescence NSCs, as well as differentiating progeny. During mitosis, unlike SOX2, MYEF2 is not retained on the mitotic chromatin indicating it is not a “bookmarking” factor. CRISPR/Cas9 mediated knock-out of Myef2 suggests that although it is not essential for the continued proliferation of GSCs, it has a role in regulating the exit from quiescence. Consistent with this, in Myef2 knock-out mouse GSCs, the rate of tumour progression is slower, and mice have a significant survival advantage, suggesting there is an important role for Myef2 in driving tumour growth. In this thesis, I also describe the use of SMASh tag degron technology to precisely control the degradation of SOX2 in mouse NSCs. SMASh, a drug degradable selfcleaving degron, was fused to the C-terminal of endogenous SOX2 in mouse NSCs. We find that this works well and can therefore be a powerful tool in future studies of GSCs.
URI
https://hdl.handle.net/1842/36925

http://dx.doi.org/10.7488/era/226
Collections
  • Edinburgh Medical School thesis and dissertation collection

Library & University Collections HomeUniversity of Edinburgh Information Services Home
Privacy & Cookies | Takedown Policy | Accessibility | Contact
Privacy & Cookies
Takedown Policy
Accessibility
Contact
feed RSS Feeds

RSS Feed not available for this page

 

 

All of ERACommunities & CollectionsBy Issue DateAuthorsTitlesSubjectsPublication TypeSponsorSupervisorsThis CollectionBy Issue DateAuthorsTitlesSubjectsPublication TypeSponsorSupervisors
LoginRegister

Library & University Collections HomeUniversity of Edinburgh Information Services Home
Privacy & Cookies | Takedown Policy | Accessibility | Contact
Privacy & Cookies
Takedown Policy
Accessibility
Contact
feed RSS Feeds

RSS Feed not available for this page