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Click here to search FindAPhD.com for PhD studentship opportunitiesAbout the Project
The first aim of this studentship is to identify regulators of insulin binding, uptake and nuclear import using genome wide CRISPR screens with fluorescently-labelled insulin. Preliminary data show that the fluorescently labelled insulin rapidly accumulates in the nucleus of hepatoma cells. To accomplish this aim the prospective student will learn to generate stable cells lines, FACS cells, perform DNA sequencing and analyse CRISPR screening data. Following identification of genes that alter insulin handling, these effects will be validated using siRNA in primary human hepatocytes with live cell imaging, immunofluorescence and classical biochemistry cell fractionation techniques. Lastly, the impact of altered insulin handling will be assessed by measurement of protein phosphorylation, metabolic flux and gene expression in insulin stimulated primary human hepatocytes with validated genes knocked down.
Successful completion of this project will identify novel factors involved in insulin binding, uptake and import into the nucleus in hepatocytes and determine how altered insulin handling influences downstream responses. This research will largely be performed at the Novo Nordisk Research Centre Oxford which houses the equipment and expertise necessary to perform genome-wide CRISPR screens. Additional validation studies will be performed at the Department of Genetics, Anatomy and Physiology.
Attributes of suitable applicants:
Ideal candidates for this PhD studentship will have strong academic background (1st or 2.1 BSc) in a relevant scientific subject. Organisational skills, personal drive and ability to work well with others are desired in a suitable candidate.
This project is supported through the Oxford Interdisciplinary Bioscience Doctoral Training Partnership (DTP) studentship programme. The student recruited to this project will join a cohort of students enrolled in the DTP’s interdisciplinary training programme, and will be able to take full advantage of the training and networking opportunities available through the DTP. For further, details please visit www.biodtp.ox.ac.uk.
Funding Notes
References
Najjar SM, Perdomo G. Hepatic Insulin Clearance: Mechanism and Physiology. Physiology (Bethesda). 2019;34(3):198‐215. doi:10.1152/physiol.00048.2018