Identification of Alternative Transcript Isoform Switches in Ageing and Senescene using Next Generation Sequencing (RNA-seq) (360G-Wellcome-202556_Z_16_Z)

During my project I aim to identify genes with alternatively expressed isoforms in mice when the individuals are subject to normal ageing, and/or under calorie restricted diet ageing. I will further study the selected candidate isoform switches, such as those predicted to regulate insulin signalling for mTOR pathways, in depth, with the intention of understanding their mechanism of regulation and how this causes the debilitating effects as so commonly associated with ageing. As the calorie-restricted diet has proved to significantly increase lifespans of mice I am personally interested to find isoforms and the mechanism that results in this drastic change between the two populations. Using the results of the bioinformatics analysis I will select candidate splice switches through filtering the data based on: statistical measure; relevance to ageing; relevance to restricted diet ageing. Once identified, the isoform switching regulatory mechanism will be studied using mini-gene constructs and designed primers to confirm the alternative events across ageing from mouse RNAs. In the future site directed mutagenesis will be used to identify the regulatory elements involved. These results could be used to target regulation of specific isoforms that may play roles in causing age-related cellular loss of function and cell senescence.