Project 1: Investigating the possible interactions between the meis1 and onecut1 proteins with adjacent LincRNAs and Project 2: Investigating hypoxia inducible factor transcriptional activity in the liver

Frampton, Nicholas Ross (2014). Project 1: Investigating the possible interactions between the meis1 and onecut1 proteins with adjacent LincRNAs and Project 2: Investigating hypoxia inducible factor transcriptional activity in the liver. University of Birmingham. M.Res.

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Abstract

Project 1 LincRNAs are molecules transcribed from regions with no protein coding function, which may regulate nearby proteins. Linc_meis1 and Linc_Onecut1 are LincRNAs located adjacent to two well-studied proteins Meis1 and Onecut1 and are the focus of this project. Analysis using whole mount in situ hybridisations attempted to visualise the native expression patterns of each LincRNA within a zebrafish model. Anti sense oligonucleotide morpholino knockdowns attempted to determine the importance of each LincRNA for adjacent protein expression and study any generated phenotypes. Results have been inconclusive; experiments are on going. A better understanding of these RNA molecules may improve understanding of developmental.
Project 2 Hypoxia inducible factors (HIFs) are transcription factors involved in regulation of stress responses associated with liver disease. The aim of this investigation was to study HIF2α stabilisation and activity within hepatocytes after exposure to hypoxia and the pro-inflammatory cytokine TNFα. Using a hypoxia responsive element (HRE) luciferase reporter gene assay, it was determined that exposure to low oxygen stabilised HIFs and stimulated HRE transcriptional activity. HRE transcriptional activity differed under two different hypoxic conditions (1% versus 3% oxygen; P<0.05, unpaired t-test). Hypoxic responses were compared in DMSO-differentiated and naïve Huh-7.5 cells with no significant difference, but a change in activity followed chronic hypoxic exposure. This kinetic data could contribute towards understanding of HIF regulation.

Type of Work:

Thesis (Masters by Research > M.Res.)

Award Type:

Masters by Research > M.Res.

Supervisor(s):