Role of m⁶A RNA "reader" YTHDF2 in Mammalian Germ Cells and Early Embryo Development

RNA modifications affect the processing and metabolism of mRNA which ultimately leads to changes in gene expression. Regulation of gene expression is essential for every tissue and every organism, specifically in cell fate determination. N6-methyladenosine (m6A) is one of the most abundant and conserved internal mRNA modifications that plays important role in regulation of gene expression in development and disease. The m6A-RNA modification mostly destabilizes mRNA levels and its role is mediated by protein that binds the m6A-RNA with high affinity, YTH-domain family protein 2 (YTHDF2). However, little is known about the role of m6A-RNA modification in vivo and YTHDF2 mechanism of regulation. To have a comprehensive understanding of m6A-RNA role in vivo we generated a state of the art complex tag and conditional allele of Ythdf2. We used this allele to develop intricate biochemistry technique for the identification of YTHDF2 targets. Furthermore, we show preliminary results on its protein partners. To better understand the physiological relevance of YTHDF2 and m6ARNA function we used the complex tag and conditional allele to generate a YTHDF2 deficient mice. Here we show that deletion of Ythdf2 is partially permissive and results in female infertility. Through conditional mutagenesis we demonstrate the importance of YTHDF2 in the generation of competent oocytes that can support early embryo development. Our molecular and bioinformatics analysis show that YTHDF2 is important for the fine tuning of the maternal transcriptome through destabilization and that its function is dependent on m6A-RNA modification.

Name dc:type.qualificationname

Doctor of Philosophy

Level dc:type.qualificationlevel

Doctoral Thesis

Grantor dc:publisher.institution

University of Dundee

Year dc:date.issued

2017

Author dc:creator

• Ivanova, Ivayla

Advisors dc:contributor.advisor

• O'Carroll, Dónal
• Keyse, Stephen
• Aulehla, Alexander
• Enright, Anton

• RNA modification
• germ cells
• RNA binding protein
• RNA destabilization
• Early embryo development

Language dc:language

eng

Identifier

oai:discovery.dundee.ac.uk:studenttheses/b89c4262-33f0-4809-908b-3f7b0d21d6a5

OAI identifier oai:identifier

oai:discovery.dundee.ac.uk:studenttheses/b89c4262-33f0-4809-908b-3f7b0d21d6a5