Missions

Mycobacterium abscessus is an emerging non-tuberculous mycobacterium (NTM) responsible for severe pulmonary infections, particularly in patients with cystic fibrosis. Treating these infections remains extremely challenging due to the bacterium's resistance to most classes of antibiotics. We are focused on identifying new surface receptors belonging to the tetraspanin family that facilitate the internalization of mycobacteria by macrophages. Our original research indicates that M. abscessus utilizes various tetraspanins to enter and invade macrophages during the early stages of colonization.

The proposed project will take a multidisciplinary approach to investigate, at the cellular and molecular levels, the role of tetraspanins CD9 and CD63, as well as their bacterial ligands, in M. abscessus infection. The outcomes of this project are expected to improve our understanding of the early events in the infectious process. Furthermore, this study could lead to a novel and innovative therapeutic approach targeting these surface receptors and their ligands, aiming to inhibit mycobacterial internalization by host cells and disrupt the infection process.

Activités

• Use biochemical techniques (production of recombinant proteins, pull-down, TURBOID, yeast two-hybrid) to identify bacterial ligands of tetraspanins CD9 and CD63.
• Clone the bacterial ligands into various vectors to determine their localization, produce them as recombinant proteins, couple them to fluorescent beads, delete them, and complement the mutants in M. abscessus.
• Apply the CRISPR/Cas9 technique to generate tetraspanin-deficient human macrophages, followed by their complementation.
• Investigate the role of bacterial ligands involved in the interaction with CD9 and CD63 during early M. abscessus infection by assessing the attachment and viability of the generated mutants in relevant models, including wild-type or tetraspanin-deficient macrophages.
• Perform fluorescence microscopy using a confocal microscope to analyze the localization of the bacterial ligands of both tetraspanins, as well as assess the ability of M. abscessus mutants to infect human macrophages and survive over time.

Compétences

• PhD in Bacteriology preferred, but feel free to apply if you have experience with other pathogens, whether parasitic or viral.
• Proficiency in molecular biology techniques, including PCR, cloning, transformation, and mutant construction by gene deletion.
• Proficiency in cellular biology techniques, with experience in cell culture, eukaryotic cell infection, and generating mutants in human cells using the CRISPR/Cas9 strategy.
• Expertise in high-resolution microscopy.
• Experience in techniques aimed at isolating protein complexes.
• Strong problem-solving skills with the ability to work both independently and collaboratively.
• Excellent written and verbal communication skills in English.

Contexte de travail

Nearly all the equipment and platforms required for the project are available in the laboratory or on-site. The position is within a sector subject to the Protection of Scientific and Technical Potential (PPST), and therefore, in accordance with regulations, your arrival must be authorized by the competent authority of the Ministry of Higher Education and Research (MESR).

Le poste se situe dans un secteur relevant de la protection du potentiel scientifique et technique (PPST), et nécessite donc, conformément à la réglementation, que votre arrivée soit autorisée par l'autorité compétente du MESR.

Contraintes et risques

Experiments and monitoring of the infection may require working outside regular hours, including on weekends.