SKM 2023 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 26: Focus Session mRNA Physics
BP 26.5: Vortrag
Donnerstag, 30. März 2023, 16:30–16:45, TOE 317
Codon position-specific engineering of translation kinetics — Judith Müller1, Gerlinde Schwake1, Anita Reiser1, Daniel Woschée1, Zahra Alirezaeizanjani3, Joachim Rädler1, and •Sophia Rudorf2 — 1Faculty of Physics, Ludwig Maximilian University of Munich — 2Faculty of Natural Sciences, Leibniz University Hannover — 3independent researcher
Recently, we introduced Live Imaging on Single Cell Arrays (LISCA), which combines time-lapse microscopy of single cells on microstructured surfaces with automated image analysis. LISCA enables us to observe and analyze the time-evolution of protein expression in hundreds of individual cells in parallel. Here, we combine LISCA with our software OCTOPOS, which simulates ribosome movement on the ORF and is used to generate reporter genes with varying ribosome speeds and densities. We predict and monitor the translation kinetics of synonymous variants of eGFP to determine mRNA functional lifetimes and translation rates with high accuracy in single lung tissue cells. Our approach allows us to study how provoked ribosome jams on specific positions within the ORF influence mRNA stability, thus linking ribosome dynamics and mRNA biophysics.