Quantitative comparison of methodologies for translation site imaging in living cells

RNA 2026 Open access

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Authors: Agata D. Misiaszek, Esther Griesbach, Egle Jaugaite, Aurelio Ortale, Jan Eglinger, Tobias Hochstoeger, Jeffrey Alan Chao
DOI: 10.1261/rna.080881.125
Cited by: 0
References: 32

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 single-chain variable fragment or 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.

Fields of Study & Subjects

RNA and protein synthesis mechanisms Monoclonal and Polyclonal Antibodies Research Click Chemistry and Applications

Publication details

Journal / source
RNA
Volume
32
Issue
9
Pages
1355–1364
Publication date
16 June 2026
Language
EN