Genetic screens of RNA binding proteins identify a role for N6-methyladenosine in promoting B cell differentiation

Post-transcriptional regulation of gene expression is mediated in part by RNA binding proteins (RBPs). Roles for RBPs have emerged in the immune system but remain largely unexplored. My project utilised high-throughput genetic screens to identify roles for RBPs in lymphocytes. Initially, I generated custom sgRNA libraries targeting RBPs to facilitate CRISPR/Cas9 mediated gene knockout screens in either human or mouse contexts. Then, I employed these reagents to identify roles for RBPs in redox signalling, B cell lymphomagenesis, CD8 T cell activation, and B cell terminal differentiation. These identified over a dozen potential avenues of future investigation into the roles of RBPs in lymphocytes. I chose to further investigate the role of the RNA modification N6-methyladenosine (m6A) in B cells. This work demonstrated a role for the m6A binding protein Ythdf2 in promoting B cell terminal differentiation, independently of a role in proliferation or survival. In the future, I hope to validate an in vivo role for Ythdf2; and further elucidate the in vitro molecular mechanism.

Name dc:type.qualificationname

Doctor of Philosophy (PhD)

Level dc:type.qualificationlevel

Doctoral

Grantor dc:publisher.institution

University of Cambridge

Year dc:date.issued

2020

Author dc:creator

• Turner, David

Advisors dc:contributor.advisor

• Turner, Martin
• Hodson, Daniel

• RNA binding proteins
• CRISPR/Cas9
• m6A
• YTHDF2

Licence

• https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/1c1da034-5aef-412d-b4d9-7a6cb8ef3bc7/download
• https://www.rioxx.net/licenses/all-rights-reserved/

Language dc:language

eng

Author Identifier

0000-0001-6225-2033

OAI identifier oai:identifier

oai:www.repository.cam.ac.uk:1810/316125