Jia-Ying Su

Abstract In eukaryotic cells, reactive oxygen species (ROS) serve as crucial signaling components. ROS are potentially toxic, so constant adjustments are needed to maintain cellular health. Here we describe a single-cell, mass cytometry-based method that we call signaling network under redox stress profiling (SN-ROP) to monitor dynamic changes in redox-related pathways during redox stress.

Quality control Request a detailed protocol We downloaded Perturb-seq datasets of retinal pigment epithelial cells from the RPE-1 cell line, and myeloid leukemia cells from the K562 cell line (Replogle et al., 2022). We used the genome-wide dataset version for the latter. We downloaded the datasets from the scPerturb database on Zenodo (Green, 2022) with the same quality controls as the original paper. Replogle et al.

eLife Assessment This valuable study combines massively parallel reporter assays and regression analysis to identify sequence features in untranslated regions contributing to the stability of in vitro transcribed mRNA delivered to cells. The strength of evidence presented is solid, although some points about half-life measurements and the relevance of identified sequence features to native transcript stability will inform future discussion surrounding the present study.