Quantitative description of protein interactions in the Wnt signalling network in vivo. Biosciences, PhD (GW4 BioMed MRC DTP....

College of Life and Environmental Sciences

University of Exeter

Quantitative description of protein interactions in the Wnt signalling network in vivo. Biosciences, PhD (GW4 BioMed MRC DTP) Ref: 3227

About the award
Supervisors

Lead Supervisor:
Dr Steffen Scholpp, Biosciences, Living Systems Institute, University of Exeter

Additional Supervisors:
Professor Paul Verkade, University of Bristol
Professor Robert Kelsh, University of Bath
Professor Christian Soeller, Physics, Living Systems Institute, University of Exeter

Project Details

Signalling molecules coordinate development and tissue homeostasis of multicellular organisms. Abnormal regulation of these signalling events leads to a diverse array of devastating diseases including cancer. The family of Wnt signalling proteins regulate vertebrate development by providing both quantitative and directional information. Furthermore, upregulation of the production of Wnt ligands is observed in many cancers leading to uncontrolled growth of tumours. Recently, we broke new ground by showing that Wnt proteins are transported by signaling filopodia - called cytonemes - in fish, mouse and human (Nat Comms, 2015, eLife, 2018). A quantitative description of cytonemal Wnt transport is still lacking. However, this knowledge is a prerequisite to understand and control this signalling pathway. We are now in the exciting position to overcome this challenge. In this project, we will generate transgenic knock-in zebrafish line by genome editing with various fluorescent proteins (XFPs) in endogenous loci in collab. with R Kelsh, Bath. To complement our approach, we will knock-in smaller tags (i.e. HA) to visualize ligand distribution in fixed tissue in combination with generation of Tol2-based transgenic GFP-fusion zebrafish lines based on characterized promoter sequences i.e. for dynamic aspects. Next, we will visualize ligand-receptor interactions with two imaging methods based on fluorescence correlation spectroscopy (FCS) and by electron microscopy. We showed that an in-vivo FCS technique can be used to characterize protein mobility and quantify binding affinities in the living zebrafish (Nature, 2009; eLife, 2018).

In this project, we will characterize the motion of Wnt molecules, quantify these on the cytonemes and measure their interaction with Fzd receptors in the zebrafish and in gastric cancer cells. We will complement our approach with a detailed CLEM approach. In collaboration with P Verkade, Bristol, we will use the APEX-GBP approach (Ariotti et al., 2015 & EMBO CLEM course in Bristol by R. Parton) to achieve fast, sensitive, and reliable localization of GFP-tagged Wnt proteins to nanometer resolution. Together with the biophysist group of C Soeller, we have the possibility to complement our analysis with FLIM-FRET. The aim of this project is the quantitative description of Wnt trafficking in vertebrate tissue with the goal to regulate Wnt trafficking in diseased tissue such as cancer.

The student selected for this project will develop invaluable skill sets in both experimental genetics, and various high-end microscopy techniques whilst also making a significant contribution to the understanding of signalling biology in gastric cancer. These mixed skill sets will make the candidate a highly desirable recruitment prospect for future academic and industrial employers. The student will be based at the LSI, UoExeter, but will spend time at the UoBristol learning imaging techniques and at the UoBath to generate reagents.

Eligibility and Residency Requirements
To be eligible for a full award (fees and stipend) from a Research Council, a UK or EU student must have no restrictions on how long they can stay in the UK and have been ordinarily resident in the UK for at least 3 years prior to the start of the studentship.

An EU student who has not been resident in the UK for at least 3 years prior to the start of the studentship is generally eligible for a fees-only award from the Research Council: to be eligible for a fees-only award, a student must be ordinarily resident in a member state of the EU; in the same way as UK students must be ordinarily resident in the UK.

For our regular recruiting round, the Doctoral Training Partnership has additional funding from the partner universities to support a limited number of EU students who do not meet the UK residency requirements, so long as they meet the criteria for a fees-only award: these studentship will therefore be fully funded (fees and stipend).

For full details on eligibility, please refer to the MRC GW4 BioMed website

Students with 'International' status are unfortunately not eligible to apply.

Entry requirements

Academic criteria
In addition to those with traditional biomedical or psychology backgrounds, the DTP welcomes students from non-medical backgrounds, especially in areas of computing, mathematics and the physical sciences, and can fund additional training, including Masters to assist discipline conversion. Applicants for a studentship must have obtained, or be about to obtain, a First or Upper Second Class UK Honours degree, or the equivalent qualifications gained outside the UK, in an area appropriate to the skills requirements of the project. Applicants with a Lower Second Class degree will be considered if they also have a Master’s degree or have significant relevant non-academic experience.

English requirements
If English is not your first language you will need to have achieved at least 6.5 in IELTS (and no less than 6.5 in any section) by the start of the programme.

General funding sources

For general sources of funding that may apply to this study opportunity, visit the page(s) below.

Funding information

Funding applies to:
EU applicants (including UK)
Funding notes:

This studentship is funded through GW4 BioMed MRC Doctoral Training Partnership. It consists of full UK/EU tuition fees, as well as a Doctoral Stipend matching UK Research Council National Minimum (£14,777 for 2018/19, updated each year) for 3.5 years.

For further information relating to the funding please see the main MRC GW4 BioMed website

This project is in competition with 69 other projects; up to 18 studentships will be available.

Contacts and how to apply

Administrative contact and how to apply:

For further information and details of how to apply please see here

Application deadline:

23 November 2018