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> Stages de M2 > Liste des stages proposés pour l’année 2020-2021 > Identify new players involved in sperm mitochondria clearance

Identify new players involved in sperm mitochondria clearance

proposé par Vincent Galy, UMR7622 - Laboratoire de Biologie du Développement, Institut de Biologie Paris Seine, Sorbonne Université, Paris

Projet de stage : After fertilization, the autophagy machinery selectively recognizes and eliminates the sperm mitochondria. One possible mechanism underlying the recognition of the sperm mitochondria is that sperm mitochondria expose a specific mark. In order to identify this mark, we will test the effect of RNAi depletion of candidate mitochondrial proteins on sperm-mitochondria stability in the embryo. We generated a list of 10 candidates mitochondrial proteins. The goal of the project will be to trigger their RNAi depletion from spermatozoa and evaluate the impact on sperm mitochondria fate in the embryos produced. This will be done by methods classically used in the lab including PCR assay, live imaging and immuno-staining. The most interesting candidates will be further analysed by GFP translational reporters generated by CRISPR-Cas9 genome editing. This work will contribute to elucidate the molecular mechanisms of sperm mitochondria clearance. Techniques mises en œuvre par le stagiaire : RNAi, live imaging, immuno-fluorescence, PCR, CRISPR-Cas9 genome editing

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