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Regensburg 2022 – scientific programme

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BP: Fachverband Biologische Physik

BP 12: Poster 2

BP 12.55: Poster

Tuesday, September 6, 2022, 17:30–19:30, P4

Live imaging on single cell arrays (LISCA) as platform to study mRNA codon optimization based on ribosome modelling — •Judith Müller1, Gerlinde Schwake1, Anita Reiser1, Daniel Woschée1, Zahara Alirezaeizanjani3, Joachim Rädler1, and Sophia Rudorf21Ludwig-Maximilians-Universität, München — 2Leibniz Universität, Hannover — 3Max Planck Institute of Colloids and Interfaces, Potsdam

mRNA based therapies have the potential to evolve as one of the most powerful therapeutic technologies of our future. Massive efforts have been made to deeply study the underlying mechanisms of mRNA delivery and translation. Synonymous re-coding of the mRNA's open reading frame is one approach to investigate and optimize the physics of mRNA translation. In this project, we evaluate the potential of bias in codon usage on influencing the mRNA's translation and degradation kinetics. Live imaging on single cell arrays (LISCA) enables the quantification of translation of hundreds of single cells in parallel on microstructured surfaces. By describing the translation in biochemical rate equations, we analyse mRNA expression and degradation rates with high accuracy. Ribosome movement on the open reading frame (ORF) is simulated to generate mRNA constructs coding for reporter genes with varying ribosome speeds and densities. We observe distinct differences in expression and degradation rates for GFP mRNAs with various optimized ORFs in agreement with simulation. Secondly, we study how specifically provoked ribosome jams on the ORF influence mRNA stability.

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