Noncanonical amino acid incorporation enables minimally disruptive labeling of stress granule and TDP-43 proteinopathy

  • bioRxiv. 2025 Oct 17:2025.10.17.683020. doi: 10.1101/2025.10.17.683020.
Hao Chen  1  2 ,  Haocheng Wang  1 ,  Yuning Lu  1  2 ,  Peng Chen  1  2 ,  Zhongfan Zheng  1  2 ,  Tao Zhang  1  2 ,  Jiou Wang  1  2
Affiliations
  • 1. Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, 21205, USA.
  • 2. Department of Neuroscience, School of Medicine, Johns Hopkins University, Baltimore, MD, 21205, USA.
PMID: 41279779 DOI: 10.1101/2025.10.17.683020
Abstract

We report a minimally disruptive labeling strategy for stress granule protein G3BP1 and ALS-linked protein TDP-43 using the fluorescent noncanonical amino acid Anap. By integrating genetic code expansion with rational site selection, we achieved precise incorporation of Anap that preserves protein structure and function. In live cells and neurons, Anap labeling faithfully recapitulated localization, stress-induced dynamics, and recovery behavior, outperforming conventional fluorescent tags and enabling physiologically relevant visualization of protein pathobiology.

Keywords
Anap labeling; Genetic code expansion; Stress granule; TDP-43.
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