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bioRxiv · 10.1101/2025.10.29.685314

Quantitative comparison of methodologies for translation site imaging in living cells

Misiaszek, A. D.·Griesbach, E.·Jaugaite, E.·Ortale, A.·Eglinger, J.·Hochstoeger, T.·Chao, J. A.
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Abstract

Single-molecule imaging of translation sites in living cells has enabled the dynamics of protein synthesis to be investigated with high spatial and temporal resolution. These methodologies utilize the interaction between a multimerized epitope tag and its cognate fluorescent nanobody to detect nascent polypeptides as they emerge from the ribosome. Here, we present a systematic comparison of current methodologies and determine that the ALFA-tag reduces perturbations of mRNA expression and increases the fluorescent signal of translation sites.

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Misiaszek, A. D., Griesbach, E., Jaugaite, E., Ortale, A., Eglinger, J., Hochstoeger, T., Chao, J. A.. 2025-10-30. Quantitative comparison of methodologies for translation site imaging in living cells. https://doi.org/10.1101/2025.10.29.685314

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