Partial human genome RNAi screen to identify genes involved in brain size regulation and cancer development.

Project description

The most common cause of autosomal recessive primary microcephaly (MCPH), a neurodevelopmental disorder characterized by reduced brain size and mental retardation, is mutation in the Abnormal Spindle Microcephaly Assembly (ASPM) gene  (formerly named Abnormal Spindle Microtubule Associated gene)(1). ASPM is localized at the spindle poles during mitosis. ASPM knockdown using small interfering RNAs (siRNAs) results in reorientation of the mitotic spindle/cleavage furrow and cytokinesis failure (2). ASPM is an extremely important protein as changes in cell division orientation during neurodevelopment has serious consequences on the size of the developing brain and spindle misorientation results in cancer development through induction of genomic instability, tissue disorganisation, metastasis and expansion of cancer stem cells.

Reverse genetic screens using libraries of small interfering RNAs are a powerful tool in identifying multiple genes involved in developmental and disease pathways (3). To identify genes involved in brain size regulation and cancer formation two siRNA screens (human protein kinase and human ubiquitin libraries) have been performed using loss of ASPM at the spindle pole as the screen readout. The student will (i) analyse the screen results to identify genes involved in ASPM regulation and localisation, mitotic progression and division orientation and cellular cytotoxicity (ii) perform siRNA knockdown experiments to validate candidate gene hits (iii) perform molecular and cell biology studies to further characterise the nature of the hits, how they modify or interact with ASPM and what is the specific function of the interaction in relation to cell division, to identify candidate MCPH genes and identify novel cancer therapeutic targets.

Timeliness and training opportunities:

Cell division is one of the most fundamental cellular processes and this project aims to further our understanding of the regulation of mitosis with specific reference to neurogenesis and cancer formation. The project will provide interdisciplinary training in functional genomics approaches (comprising high-throughput screening and image analysis) followed by molecular and cell biology studies (immunofluorescence microscopy, Western blotting, Co-Immunoprecipitation studies, cloning, and specific functional assays based on the nature of the candidate hits). There will be an early emphasis on image analysis and the interpretation of large datasets, which will provide important transferable bio-analytical and bioinformatics skills.

References:

• Bond et al., (2002), Nat Genet. 32:316-20. PMID: 12355089
• Higgins et al., (2010) BMC Cell Biol 11:85.  PMID: 21044324
• Wheyway et al., (2015) Nat Cell Biol. 17:1074-1087 PMID:26167768

Entry requirements

This project is available immediately to both Home/EU rate applicants and International applicants who are able to self-fund their studies. Students must be able to provide the appropriate level of fees based on their fee status plus laboratory consumables costs per year. This is in addition to the provision of personal living expenses.

You should hold a first degree equivalent to at least a UK upper second class honours degree in a relevant subject.

Candidates whose first language is not English must provide evidence that their English language is sufficient to meet the specific demands of their study, the Faculty minimum requirements are:

• British Council IELTS - score of 6.5 overall, with no element less than 6.
•  TOEFL iBT - overall score of 92 with the listening and reading element no less than 21, writing element no less than 22 and the speaking element no less than 23.

Applicants with sufficient funding must still undergo formal interview prior to acceptance in order to demonstrate scientific aptitude and English language capability.

How to apply

Applications can be made at any time. Potential applicants are welcome to contact Dr Jacquelyn Bond with informal enquiries about this research project.

To formally apply for this project applicants should complete a Faculty Scholarship Application form and send this alongside a full academic CV, degree transcripts (or marks so far if still studying) and degree certificates to the Faculty Graduate School. 

We also require 2 academic references to support your application. Please ask your referees to send these references on your behalf, directly by email.

If you have already applied for other scholarships using the Faculty Scholarship Application form you do not need to complete this form again. Instead you should email to inform us you would like to be considered for this scholarship project.