CRISPR CAS9 Genome Editing to Study PIP5K1A in Cancer Cells

Human PIP5K1A is one of a family of kinases that generate the signalling molecule phosphatidylinositol 4,5-bisphosphate (PIP2) in cells. This membrane associated lipid and its derivatives have diverse roles in endocytosis, cell movement and receptor-mediated signalling, e.g. via the Wnt signalling pathway. Consistent with this, PIP5K1A shows high expression in tumours (Sarwar et al., Oncotarget 2016), suggesting it contributes to malignant cancer by enabling metastasis.

Research from our collaborators in Sweden, suggests a key role for PIP5K1A in malignant prostate cancer that can be targeted by drug intervention, as shown in xenograft models (Semenas et al., PNAS 2014). The data also highlight a previously unknown intersection of phosphoinositide and androgen signalling pathways, demonstrating the potential of combined therapies as effect treatments in animal models (Sarwar et al., Oncotarget 2016). However, little is known regarding the molecular interactions of PIP5K1A underlying these functions, or its effects on gene expression in cancer cells.

The goals of this project are to investigate the structure and function of human PIP5K1A. A combination of techniques including CRISPR CAS9 genome editing, RNA Seq and functional assays will be employed to determine how loss or overexpression of PIP5K1A alters gene expression in prostate cancer cells. Rescue experiments will be employed in PIP5K1A deficient cell lines, to explore function. We are also working towards recombinant expression studies in E.coli towards defining the crystal structure of human PIP5K1A.

Environment
The project will be carried out within the Gene Regulation & RNA Biology Group (circa 30 researchers) within the Division of Molecular & Cellular Sciences. The School of Pharmacy provides a vibrant cross-disciplinary research environment, with excellent support and training facilities for PG students.