Translation inhibition with CHX in mouse embryonic stem cells

Gene-expression noise can influence cell-fate choices across pathology and physiology. However, a crucial question persists: do regulatory proteins or pathways exist that control noise independently of mean expression levels? Our integrative approach, combining single-cell RNA sequencing with proteomics and regulator enrichment analysis, reveals 32 putative noise regulators. The approach utilizes global translation inhibition (i.e., potential protein regulators), and single-cell RNA sequencing (scRNA-seq) to quantify the changes in noise of all transcripts (i.e., potential mRNA targets). This dataset corresponds to the aforementioned scRNA-seq experiment upon translation inhibition with cycloheximide. Mouse embryonic stem cells (mESCs) were seeded and growth in serum/LIF conditions. 24 hours post-seeding, cells were treated with cycloheximide for 3 or 6 hours, or EtOH as control. After treatment cells were dechached and frozen in preparation for single-cell RNA sequencing. Cells were shipped in dry ice and single-cell RNA sequencing was performed from a freshly thawed sample, through single-cel barcoding (10x Genomics'Chromium) and sequencing (NextSeq 2000, Single Cell 3'v3 chemistry).