Rio-seq Analysis on SETD2-Deficient and Control Intestinal Epithelial Cells

Protein abundance does not necessarily correlate with mRNA levels, highlighting the highly variable efficiency of translation. Ribosome biogenesis, the process of ribosome production, affects translation efficiency and plays a crucial role in various biological processes. However, its role in maintaining epithelial barrier integrity has not been definitively established. SETD2 is a methyltransferase critical for the integrity of epithelial barrier. Here, we examined intact intestinal barrier with SETD2 present and compromised intestinal barrier with SETD2 loss. Since mRNA translation in ribosomes determines protein abundance and discordant translational efficiency (TE) causes protein-mRNA discrepancies, we utilized ribosome profiling sequencing (Ribo-seq) to investigate genome-wide TE changes during the loss of epithelial integrity. In this process, mRNA undergoing translation, represented as ribosome footprints (RFs), was sequenced and compared to total mRNA sequencing data to illustrate changes in the TE of genes.