We are recruiting an exceptional individual to join as a Research Technician in the laboratory of Prof Gopal Sapkota. This is a fixed-term appointment for 6 months.

We are looking to identify substrates of protein phosphatase PPP3CA and PPP3CB in human cells. This project aims to generate cell lines in which PPP3CA and PPP3CB will be knocked in with dTAG and bromoTAG degrons, respectively, using CRISPR/Cas9 genome editing. This will allow us to acutely and simultaneously degrade PPP3CA and PPP3CB proteins by PROTACs directed at dTAG and bromoTAG. At the same time, the project also aims to generate PPP3CA and PPP3CB gene knockouts. It is also hoped that the candidate will then be able to take some of these cells and undertake an unbiased phospho-proteomic analysis, with the help of MRC PPU mass spectrometry service, to identify PPP3CA and PPP3CB substrates.

Your priorities will include:

• Identification of cell lines to be used for CRISPR/Cas9 genome editing based on bioinformatic data available on expression of PPP3CA and PPP3CB isoforms.
• Design and perform CRSIPR/Cas9 knock-in and knockout experiments.
• Validate the CRISPR/Cas9 clones via immunoblots, PCR and DNA sequencing
• Evaluate degradation of PPP3CA and PPP3CB proteins by using PROTACs directed at dTAG and bromoTAG and determine optimal conditions.
• Undertake total and phospho-proteomic analysis to evaluate specificity and consequences of PPP3CA and PPP3CB degradation.
• Collate data and analyse.
• Dissemination of findings at regular lab meetings.

Candidate requirements:

• Hold a BSc and/or MSc in Cell Biology, Molecular Biology, Biochemistry, Proteomics or a related discipline with outstanding academic track record.
• Have a strong background of working in a research lab
• Have knowledge and experience of generating knockout and knockin cells using CRISPR/Cas9 genome editing
• Have a strong interest in signal transduction research and understand PPP3CA/B biology
• Have a strong background in molecular biology and biochemistry.
• Knowledge of proteomics is desirable.
• Have a strong ability to work independently but with excellent ability to work in a team, and an open and collaborative approach to science.
• Highly organised, motivated and meticulous, with an ability to work independently and to drive a project forward robustly and at pace.
• Have excellent communication skills and knowledge of the English language are essential.

We are one of the UK's leading universities, internationally recognised for our expertise across a range of disciplines and research breakthroughs in multiple areas, including science, medicine and engineering, amongst many others. Our purpose is to transform lives, locally and globally, which we do as a community of staff (Professional Services and academic Schools), students and alumni. Professional Services directorates are key to delivering the University strategy and driving change across the University. We are signatory to the national Technician Commitment (https://www.dundee.ac.uk/events/history-pitfalls-and-hallmarks-success-technicians-commitment).

For further information about this position please contact Prof Gopal Sapkota at g.sapkota@dundee.ac.uk. To find out more about MRC PPU please visit https://www.ppu.mrc.ac.uk/

The diversity of our staff and students helps to make the University of Dundee a UK university of choice for undergraduate, postgraduate and distance learning. Family friendly policies, staff networks for BME, Disabled and LGBT staff, membership of Athena SWAN, the ECU Race Equality Charter and Stonewall as well a full range of disability services, create an enjoyable and inclusive place to work.