A novel miRNA cluster within the circadian clock gene NPAS2 and the implications of rs1811399, an autism enriched single nucleotide polymorphism

Abstract

This PhD investigated the role of the rs181399 single nucleotide polymorphism (A or C) in the maturation of a previously unknown microRNA. microRNA are potent regulators of gene expression and their expression is finely controlled. Canonically the microRNA processing machinery recognise hairpin-loops of single stranded RNA as a substrate for processing. Mutations, such as rs1811399 can disturb the hairpin leading to reduction in expression. The work presented in this thesis demonstrates that a novel microRNA cluster is located within intron 1 of NPAS2 which is independently transcribed of its host gene. Secondly, plasmid constructs were used to integrate versions of the novel microRNA hairpin containing either an A allele or a C into cell lines. Utilising this method the impact of the C allele was noted to be deleterious to microRNA maturation. RNA protection assays have demonstrated that both precursor microRNA and mature microRNA is constitutively expressed within a multitude of tissue types, seemingly independently of its host gene. The potential impact of the C allele on genetic regulation was analysed bioinformatically by analysing potential gene targets and the pathways they participate in.

Date of Award	3 Dec 2014
Original language	English
Awarding Institution	Bangor University
Sponsors	Knowledge Economy Skills Scholarship (KESS)
Supervisor	Thomas Caspari (Supervisor)