For master students at UniGE, the lab offers several possible master projects in which you can learn a wide range of techniques. Projects are typically multi-disciplinary, including techniques such as molecular cloning, molecular genetics, live-cell imaging, correlative light-electron microscopy, or biochemical methods. Below are a couple of examples. If you are interested in discovering more, come and talk to us!

Establishing an optogenetic method of protein trapping in fission yeast 

Identifying protein function often relies on the ability to genetically or pharmacologically modulate activity, yielding phenotypes to provide clues to physiological function. However, for most proteins, small pharmacological compounds are unavailable, and genetic manipulation can be difficult. For instance, for proteins with essential roles in cellular life, deletion is not possible, and precise spatiotemporally manipulation of protein activity is required to obtain functional information. Optogenetic techniques, where light is used to induce protein-protein interaction and modify protein function, can provide such precise temporal and spatial control in single cells. The master student will establish a method of protein trapping by optogenetics in fission yeast cells, based on a published strategy (named LARIAT, for Light-Activated Reversible Inhibition by Assembled Trap; Lee et al, Nature Methods 2014). The lab has expertise with optogenetics, and we already have proof-of-principle that the method can work, but experimental conditions need to be optimized to make the method useful in many research applications. In this internship, the student will learn to use molecular genetics, cloning, yeast transformations, live-cell imaging by fluorescence microscopy, including optogenetics, and image analysis techniques.

MAPK signaling during sexual reproduction

During sexual reproduction, gametes fuse together to form the diploid zygote. In fission yeast cells, mating depends on pheromone signaling between partner cells, which triggers a conserved signaling pathway from a G-protein-coupled receptor (GPCR) through a MAPK signaling cascade. This GPCR-MAPK signaling is not only essential to induce the transcriptional changes that promote sexual differentiation, but also acts locally at the site of partner cell contact to promote cell-cell fusion (Dudin et al. 2016). To dissect the mechanisms of this signaling pathway, we have conducted phospho-proteomic screens, identifying candidate MAPK substrates. In this project, the master student will investigate the possible role and localization of candidate MAPK substrates, by constructing deletion and tagged strains, as well as strains in which the identified phosphorylation sites are mutated. For substrates with interesting location/phenotype, further analysis will involve in vitro work to confirm direct phosphorylation by the MAPK. In this internship, the student will learn to use molecular biology, yeast molecular genetics (including CRISPR), live-cell imaging, in vitro kinase assays.