Polyclonal CRISPR screening is an innovative non-biased genome-wide approach to identify key genetic determinants of a specific cellular phenotype. A mixed pool of cells, each with a single gene knock out is generated. These cells can be exposed to a specific environment with the resulting genetic changes identified by next generation sequencing. This approach has been successfully used to identify critical drivers involved in drug resistance. Hypoxic cancers are resistant to treatment and have increased metastatic potential arising from changes in signalling pathways such as those found in the apoptotic pathway. Hypoxia alters the cell death response by decreasing the expression of pro-apoptotic proteins and increasing the expression of anti-apoptotic proteins. These changes protect the cancer cells from treatment-induced apoptosis from radiotherapy or chemotherapy. Hypoxia modification has been used to improve the survival of patients with bladder and head & neck cancers with no increase in side-effects. Hypoxia modification works by alleviating the low oxygen tension. It does not eradicate the evolved hypoxic-resistant phenotype and despite its use chemoradioresistance remains a problem. Prostate cancer is a global problem with over 1 million cases diagnosed and over 350,000 deaths per year. Radiotherapy and chemotherapy are important treatments. We will use prostate cancer as a model for using a CRISPR screening approach to identify novel hypoxia-induced cell death regulators that can be used to improve the efficacy of local and systemic treatment.
Objectives: 1. Optimise experimental conditions for quantifying apoptosis when prostate cancer cells are exposed to different forms of hypoxia. 2. Using a polyclonal CRISPR screen, identify genomic changes after exposure to hypoxia alone by comparing the profiles of Annexin V positive and negative cells. 3. Validate targets from the CRISPR screen and test in vitro activity of existing drugs for improving radiotherapy response when compared with a known anti-apoptotic inhibitor.
We invite applications from recent graduates or final year undergraduates who have, or expect to obtain, a first or upper second class honours degree, or the overseas equivalent, in a relevant subject. A related Master’s degree would be an advantage.
For applicants whose first language is not English, you must supply an official IELTS or TOEFL transcript to support your application, or provide a date on which you will be taking a test.
For information on how to apply for this project, please visit the Faculty of Biology, Medicine and Health Doctoral Academy website (https://www.bmh.manchester.ac.uk/study/research/apply/). Informal enquiries may be made directly to the primary supervisor. On the online application form select PhD Cancer Sciences.
For international students we also offer a unique 4 year PhD programme that gives you the opportunity to undertake an accredited Teaching Certificate whilst carrying out an independent research project across a range of biological, medical and health sciences. For more information please visit www.internationalphd.manchester.ac.uk
Applications are invited from self-funded students. This project has a Band 3 fee. Details of our different fee bands can be found on our website (https://www.bmh.manchester.ac.uk/study/research/fees/). For information on how to apply for this project, please visit the Faculty of Biology, Medicine and Health Doctoral Academy website (https://www.bmh.manchester.ac.uk/study/research/apply/).
As an equal opportunities institution we welcome applicants from all sections of the community regardless of gender, ethnicity, disability, sexual orientation and transgender status. All appointments are made on merit.
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Wei L et al. Genome-wide CRISPR/Cas9 library screening identified PHGDH as a critical driver for Sorafenib resistance in HCC (2019). Nature Communications.
Watanabe S et al. Loss of KDM6A characterizes a poor prognostic subtype of human pancreatic cancer and potentiates HDAC inhibitor lethality (2018). Int. J. Cancer; 145:192–205.
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Kinoshita M et al. Cancer cells surviving hypoxia obtain hypoxia resistance and maintain anti-apoptotic potential under reoxygenation (2001). Int J Cancer; 9:322–326.
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