Development of a chick model to identify LSAMP gene expression enhancers

• Main Author: Lyons, Michael
• Published: University of Liverpool 2012
• Subjects: 571.6
• Online Access: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.632424

One of the major ways employed by eukaryotic cells to control gene expression is by the action of the regulatory sequences known as enhancers and repressors. An understanding of the spatial and temporal activation of these sequences would help our understanding of gene control. Because polymorphisms in these control regions have been associated with disease a way to rapidly identify if these polymorphisms were affecting gene expression would be a useful diagnostic tool. This thesis describes the development of a chick-based model to attempt to rapidly identify when and where enhancers are being activated. Using the ECR Browser, putative enhancer sequences are identified by cross-species comparison of the genomes to expose evolutionary conserved regions. By cloning an ECR, so identified, into a suitable reporter vector and transfecting cells in-ovo, the activation of the enhancer by the cell can be detected. Following a successful proof-of-concept run using the Hb9 gene, LSAMP, was selected as the first gene to investigate because of its role in development and its implication in mental health disorders and cancer. A number of ECRs were examined and six of these were shown to be enhancers proving that the system functioned.