Work

Two‐Color Timestamping of Gene Expression with a Chemigenetic Reporter System

Year: 2025

Type: article

Abstract: Stress response pathways rely on coordinated gene expression regulation. In the endoplasmic reticulum, the unfolded protein response plays an important role in maintaining the homeostasis of protein folding and processing. A chemigenetic reporter assay is developed that allows monitoring of BiP/GRP78 expression, a key regulator of the unfolded protein response. The system is based on the coexpression of the self-labeling protein HaloTag via an internal ribosome entry site. HaloTag allows for flexibility in labeling color as well as labeling timepoint. A two-color timestamping strategy is designed that provides improved readout sensitivity. The reporter system can be used in live-cell imaging as well as flow cytometry. It is applied in a high-content screening experiment in which a new promising activator of BiP/GRP78 expression was identified.

Source: ChemBioChem

DOI: 10.1002/cbic.202500494