Thomas Laskarzewski – NIH Trainee 2024-2025
Research Progress: I am in my second year of my PhD in the Maresca lab studying Metazoan kinetochore structure. My project involves taking a top-down approach to kinetochore structure by oligomerizing CENP-C, a disordered kinetochore protein, with the optogenetic oligomerization tag, Cry2. Optogenetic oligomerization results in CENP-C oligomers that co-localize other kinetochore proteins, resulting in kinetochore particles. We plan to perform biochemical isolation of these kinetochore particles and use them for structural (electron microscopy), functional (movement of kinetochore particles on dynamic and stabilized microtubules), and compositional analysis (mass spectrometry). In my first 6 months in the lab, I spent time cloning constructs and creating stable cell lines, then characterizing expression through western blotting and co-localizing proteins through live cell fluorescent microscopy and fixed immunofluorescence. More recently, I have begun isolation of kinetochore particles, using strep-tavidin conjugated magnetic beads to pull-down kinetochore components.