Carole Seum
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PROJECT
In Marcos Gonzalez鈥� lab, my goal is to visualize protein-protein interaction in living cells using the BIFC approach (Bimolecular Fluorescence Complementation).听This technique consists of the use of a fluorophore split into two parts and fused to two different interactors. In vivo association of these two proteins leads to the reconstitution of the fluorescence otherwise absent from each isolated fragment. The best combinations of fused moieties was first tested in Schneider-2 cells (no self-association of naked fragments) and I then observed the interaction in a living organism such as Drosophila melanogaster.听The interaction I want to evidence here is the nuclear association of pMad and Medea in the nucleus in response to Dpp signaling.
This technique will also allow me to trace in a living cell the dynamic of association of different proteins sharing a common interactor by a multicolor monitoring. This experiment can be realized using N-terminal fusions of different fluorophores and a common C-terminal part of Venus. I would like to visualize the interaction of Smox and Mad with Medea.
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education/Academic background
For 15 years I have been studying chromatin organization听and听silencing using position-effect variegation听in听Drosophila听melanogaster听as听a tool. I did听my听PhD听in听2000听in听P. Spierer鈥檚听laboratory, studying PEV using new genetic tools that were听imported into the heterochromatic compartment.听I then experienced successfully the homologous recombination technique described by Kent Golic, knocking down few genes such听as听Su(var)3-7, G9a and SETDB1, these two latest being histone听methyl transferase specific for H3听on听lysine 9, a mark known听to听be听implicated听in听gene silencing. I also created few听鈥渒nock-in鈥澨齯sing听the same technology.听I听am听now currently using CRISPR/Cas9听in听Drosophila听as听a tool for听targeted genome editing听of听many genes commonly used听in听the lab.