DNA scaffolds for tunable spatiotemporal control over receptor kinase assemblies

The INSERM U1242 laboratory is a translational research structure bringing together biologists, chemists and clinicians to better understand oncogenesis in brain and gynecological cancers. By associating biologists working on cancer stress signaling pathways, chemists developing molecular tools and cancer clinicians, the laboratory aims at deciphering how tumor cells cope with stresses to resist to death and how they could be sensitized to treatments. The candidate will join an exciting and multidisciplinary scientific environment with on-site access to cutting-edge facilities in biochemistry, biophysics, molecular biology, cell biology and structural biology. The INSERM U1242 laboratory Is affiliated to the Université of Rennes, a top French university that offers a dynamic and lively international campus with an ideal location less than 2 hours from Paris and 1 hour from Saint Malo and the beautiful Britany coast and its beaches. 

Supervisors: Dr X. Guillory (PI), C. Walter (PhD); Remuneration: 580€ net/month; Duration: 6 or 7 months between January and July 2027

Scientific background – Cellular stress induced by the abnormal accumulation of improperly folded proteins in the endoplasmic reticulum (ER) is emerging as a major actor in disease development and an appealing actionable target.[1] ER stress levels are under constant surveillance by the unfolded protein response (UPR), a major adaptive mechanism that lies at the core of cellular homeostasis and is responsible for cellular life-or-death decisions. The Inositol-requiring enzyme 1 (IRE1), the most evolutionary conserved UPR transducer, is an ER-resident transmembrane protein with a cytosolic dual kinase/RNase activity controlling pro-survival or pro-death signals. Yet, despite >20 years of investigations, the precise molecular mechanisms by which IRE1 is activated and exerts its catalytic and scaffolding functions still remain unclear and a subject of debate.[2] The group aims at deciphering the molecular mechanisms of IRE1 activation through a unique and novel prism at the interface of chemical biology, supramolecular chemistry and structural biology.

Description of the project & duties -

This internship is in support of a PhD project focusing on using a programmable synthetic platform made of DNA structures (i.e. DNA origami) to control the nanoscale organization and nature of different complexes of a kinase protein.[3]

As part of this internship, you will focus on:

1 –Expressing, purifying and functionalizing the protein kinase N-terminus using a small molecule reagent.

2 - Assembling and purifying DNA origami structures.

3 - Bioconjugating proteins on the DNA origami using DNA hybridization principles.

4- Validating by AFM microscopy the functionalized Protein-DNA nanostructure obtained.

5 – Evaluating the enzymatic activities of the protein using established biochemical assays and molecular biology techniques.

Main activities and techniques involved –

• Molecular biology (design and conception of plasmids).

• Protein expression in eukaryotic cells.

• Protein engineering, bioconjugation, purification (affinity & SEC chromatography).

• Structural characterization of protein-DNA conjugates (AFM, CryoEM).

Applicant profile – We encourage applications from highly motivated students from the national and international community with a background in chemical biology, biochemistry, biophysics, molecular biology, or other life sciences and with outstanding qualifications.