The recruited candidate will investigate the structural dynamics of green-to-red photoconvertible fluorescent proteins used in Single Molecule Localization Microscopy (SMLM) using NMR, kinetic X-ray crystallography and single-molecule imaging. The goal will be to engineer variants of enhanced photo stability to improve the quality of quantitative SMLM data.
Fluorescent proteins (FPs) are the markers of choice for many applications in advanced fluorescence microscopy. Yet, super-resolution techniques such as PhotoActivation Localization Microscopy (PALM) and single-particle-tracking PALM (sptPALM) have not yet reached maturity, notably due to the non-ideal photophysical performance of the used FP labels, in particular green-to-red photoconvertible fluorescent proteins (PCFPs). This project aims at enhancing the photostability of PCFPs by combining investigations of their structural dynamics in realistic environmental conditions with large-scale engineering and screening at the single-molecule (SM) level. The recruited candidate will concentrate on mechanistic investigations of PCFPs by NMR and crystallography. Notably, a state-of-the-art NMR toolbox has been recently developed at the Institute, specifically adapted to the study of fluorescent proteins. Applying and developing this NMR toolbox, in combination with crystallography and optical spectroscopy, will be central to the project and will be used to suggest relevant mutation sites to engineer enhanced PCFPs. The FP engineering part of the work will be performed in collaboration with our partners in Bordeaux (J.B. Sibarita, M. Sainlos), in the context of a recently awarded grant by the French “ANR”. Overall, the candidate will seek to provide key mechanistic insights into the “dark side” of PCFPs, which has remained largely overlooked thus far due to its intricate nature, pushing the knowledge of FPs beyond the state-of-the-art.
Grenoble is situated in the middle of the beautiful French Alps, and the IBS provides a unique environment for state-of-the-art integrated cellular and structural biology.
Candidates should have a strong background in biophysics. Experience in multidimensional NMR spectroscopy, X-ray protein crystallography and good knowledge of biochemistry and fluorescence microscopy will be required. Additional expertise in molecular biology, cell biology, super-resolution microscopy, single-molecule imaging and fluorescent proteins will be key advantages.
The position will be financed for 2+1 years by a recently obtained ANR grant and IBS funding already available.
Applications are now open. Please send a complete CV, a motivation letter, and at least two reference letters to Dominique Bourgeois (dominique.bourgeois@ibs.fr), before February 28, 2021.
Candidates must also feel an application at CNRS
