ATAC-seq assay of in-vitro-differentiated plasma cells

Translational control plays a crucial role in the immune system, yet the key players, molecular mechanisms, and physiological functions remain poorly understood. In this study, we performed a CRISPR/Cas9-mediated screen of translation initiation factors (TIFs) and identified eukaryotic translation initiation factor 4G2 (eIF4G2) as a major regulator of internal ribosome entry site (IRES)-mediated translation during plasma cell differentiation. Through mouse genetic studies, eCLIP-seq analysis, and CRISPR/Cas9-based functional screening of target genes, we demonstrated that eIF4G2 regulates plasma cell differentiation by binding to an IRES element in the 5' UTR of Mef2c mRNA, thereby facilitating its translation. Mef2c is a transcriptional factor. To investigate the molecular mechanisms underlying Mef2c regulation of plasma cell differentiation, we performed Mef2c CUT&Tag assay and ATAC-seq assay in in vitro differentiated plasma cells.