Nasonia vitripennis : an insect model for DNA methylation

DNA methylation is an epigenetic mechanism that regulates gene expression and chromosomal organisation. Despite being a conserved phenomena across eukaryotes, the role and location of DNA methylation differs greatly across kingdoms. In the last decade, a new field exploring the role of DNA methylati...

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Bibliographic Details
Main Author Thomas, Christian L
Format Dissertation
LanguageEnglish
Published University of Leicester 2022
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Summary:DNA methylation is an epigenetic mechanism that regulates gene expression and chromosomal organisation. Despite being a conserved phenomena across eukaryotes, the role and location of DNA methylation differs greatly across kingdoms. In the last decade, a new field exploring the role of DNA methylation in insect phenotypes has evolved. However, understanding the exact function of insect methylation has proven difficult. To elucidate this, what is needed is an insect model organism for DNA methylation studies. Nasonia vitripennis is a parasitoid wasp with a wide range of publications to its name. Unlike many insect species,Nasonia possess a fully functional DNA methylation system, displays observable methylation levels, and doesn't exhibit social behaviour. The aim of this thesis is to expand on current Nasonia literature to establish Nasonia vitripennis at the forefront of insect methylomics. I investigate DNA methylation through metamorphosis and find a dynamic methylome through developmental stages. I explore the expression profile of Nasonia DNA methyltransferase enzymes in an attempt to characterise the functionality of the expanded Nasonia repertoire and find NvDnmt1c is a female and potentially ovary specific gene. Given the links between DNA methylation and ageing in mammals I investigate the link between gene expression and DNA methylation across time in adult Nasonia. Studying whole adult females, I find little evidence of DNA methylation correlating with ageing in Nasonia vitripennis. Finally, I attempt to alter loci specific methylation in Nasonia vitripennis using a deactivated version of Cas9 fused to DNMT3a. I failed to observe transient expression of dCas9-DNMT3a, but have created a plasmid which allows easy promoter replacement for future attempts. This thesis adds to the expanding field of Nasonia methylomics, and outlines strategies to truly make this jewel wasp the dominant force in arthropod epigenetics.
Bibliography:0000000511126132
DOI:10.25392/leicester.data.21960113.v1