Oguz Can Koc

Oguz Can Koc – UMass Trainee 2023-2025

Investigating the role of CaMKIIγ in early embryogenesis and activation kinetics of CaMKII variants at different calcium oscillation frequencies. 

I have generated several plasmids for in vitro transcription and oocyte expression of different CaMKII variants (I used a FLAG-tagged T7 promotor expression system and cloned in CaMKIIα,β,γ variants with different linker lengths). I also screened various techniques to activate eggs in order to induce calcium oscillations at different frequencies. I eliminated the options of using thimerosal and caged-IP3 since they didn’t show reliable variance of oscillation frequencies we desire or had residual calcium entry following induction that created complex calcium profiles, respectively. We decided to take advantage of a photoswitchable calcium channel system where we can transiently express an engineered calcium channel with a light inducible ligand binding ability which will allow us to control the frequency of calcium oscillations in eggs. I am currently working on optimizing this system in mouse eggs. Additionally, we received the CaMKIIγ knock-out (KO) mice from our collaborators and are currently getting ready to start in-house breeding and expansion of our colony to start testing our CaMKII variants. Finally, I am testing 2 different compounds (TPEN and Cycloheximide) to activate mouse eggs in a CaMKII-independent manner which will allow us to test developmental competence of CaMKIIγ KO embryos and investigate the role of CaMKIIγ during preimplantation embryo development.