BBSRC NLD Doctoral Training Partnership: A serine proteinase inhibitor as a gatekeeper to mammalian skeletal development: from gene editing to gene evolution

This project will explore the molecular mechanisms behind a novel and essential regulator of mammalian skeletal development using cutting-edge tools to study its role in cell signalling and gene evolution across species.

Despite its central importance to vertebrate life on Earth, our understanding of the molecular events of mammalian skeletal development are not fully understood. The early skeleton is made of cartilage, ultimately becoming calcified into bone. Cartilage formation is known as ‘chondrogenesis’ - condensation of mesenchymal stem cells (MSCs) which differentiate into chondrocytes (cartilage cells) laying down a cartilage extracellular matrix (ECM). Our lab has identified a serine proteinase inhibitor (serpin) which is essential for chondrogenesis to occur. Our data support a role for this gene in driving the production of extracellular matrix (ECM), potentially through interaction with the major transcriptional regulator of chondrogenesis, SOX9. Uniquely amongst the serpin family, it also contains a DNA-binding domain which may also be important for this role.

The serpin gene architecture is also unusual. While humans and apes have a single gene copy, many animals have multiple gene paralogues (up to 14!), the relevance of which is not yet understood.

In this project, we will investigate how this protein affects stem cell differentiation into chondrocytes, and identify intracellular interaction partners using biochemistry, proteomics, and advanced single-cell microscopy techniques. The student will use gene editing (CRISPR/Cas9) to generate MSC mutant lines deficient in proteinase inhibition or DNA binding, and investigate the effect on cartilage formation. Finally, they will perform molecular phylogenetics and comparative genomics of cross-species gene evolution, inferring potential implications on mammalian skeletal development. The training for the student will be comprehensive, wide-ranging, and supported by the right expertise throughout.

The project is based in the lab of Dr David Wilkinson (Institute of Life Course and Medical Sciences; University of Liverpool). The Institute hosts a diverse research environment, with 173 PhD students (50/50% female/male, 64/36% UK/overseas) in a state-of-the-art facility (William Henry Duncan Building, est. 2016). CRISPR/Cas9 will be performed during a research stay with Professor David Young (Newcastle University), and gene evolution analysis will be carried out with support from the Professor Sebastian Shimeld (University of Oxford).

The student will join a supportive research group in a dynamic environment, garnering a range of in-demand bioscience skills in preparation for a strong future in the life sciences.

For informal enquires please contact Dr David Wilkinson (david.wilkinson@liverpool.ac.uk)

Lab website: https://www.proteolysislab.com   

Institute Website: https://www.liverpool.ac.uk/life-course-and-medical-sciences/

HOW TO APPLY:

Applications should be made by emailing bbsrcdtp@liverpool.ac.uk with:

• a CV (including contact details of at least two academic (or other relevant) referees);
• a covering letter – clearly stating your first-choice project, and optionally 2nd ranked project, as well as including whatever additional information you feel is pertinent to your application; you may wish to indicate, for example, why you are particularly interested in the selected project(s) and at the selected University;
• copies of your relevant undergraduate degree transcripts and certificates;
• a copy of your passport (photo page).

A GUIDE TO THE FORMAT REQUIRED FOR THE APPLICATION DOCUMENTS IS AVAILABLE AT https://www.nld-dtp.org.uk/how-apply. Applications not meeting these criteria may be rejected.

In addition to the above items, please email a completed copy of the Additional Details Form (as a Word document) to bbsrcdtp@liverpool.ac.uk. A blank copy of this form can be found at: https://www.nld-dtp.org.uk/how-apply.

The deadline for all applications is 12noon on Monday 22nd July 2024.

Part-Time Study Options

All NLD DTP PhDs are available as part time or full time, with part time being a minimum of 50% of full time. Please discuss potential part time arrangements with the primary supervisor before applying to the programme.

Project CASE Status

This project is not a CASE project. While individual applicant quality is our overriding criterion for selection, the NLD DTP has a commitment to fund 8 CASE projects per year - as such, CASE projects may be favoured in shortlisting applicants when candidates are otherwise deemed to be equal or a consensus on student quality cannot be reached.