Resolving the architecture of mRNA lipid nanoparticles through advanced NMR techniques

The PhD student will work on a multidisciplinary project involving the LAGEPP, Gepharm Team, and the Very High Field NMR Center of Lyon (CRMN). 

The aim of the thesis project is to contribute to the development of innovative analytical approaches to examine the internal atomic, molecular and nanoscale structure of mRNA-loaded lipid nanoparticles. The final goal is to identify the location of the mRNA cargo, probing its hydration and secondary structural elements as well as its interactions with LNP components.

Activities:

• The student will prepare tailored mRNA strand that will incorporate isotopically enriched ¹⁵N and ¹³C isotopes, and/or fluorescent nucleotides.

• The student will formulate the mRNA-based nanosystems by encapsulation or by complexation using the microfluidic techniques. The obtained particles will be characterize by standard methods such as electron microscopy to assess their size, polydispersity, and morphology

• The student will evaluate the mRNA stability in the formulation, to ensure that the developed methods are not affecting the functionality of the macromolecule.

• The student will implement high-resolution NMR experiments in the solid-state to characterize the mRNA-loaded LNPs

Skills and competences to be acquired:

Firstly, the student will learn the importance of the bibliographic research, that will be accompanied daily the practical work in the laboratory. Of key importance will be the work organization, and the ability to draft reports to track the laboratory activities. Furthermore, the student will acquire strong presentation skills in front of a heterogenous group.

The student will gain knowledge in the preparation of mRNA based on Trilink technology. He/she will then learn to use instruments dedicated at the production through microfluidic and physicochemical characterization of nanosystems, as dynamic light scattering (DLS), electron microscopy and electrophoretic light scattering (ELS) techniques, and nanoparticles tracking analysis (NTA). The student will additionally acquire basic and experimental skills in NMR spectroscopy.

To apply please send your CV and motivation letter to:

Giovanna LOLLO: giovanna.lollo@univ-lyon1.fr

Guido PINTACUDA: guido.pintacuda@ens-lyon.fr

Anne LESAGE: anne.Lesage@ens-lyon.fr