Genome-wide CRISPR library screening to identify genes involved in the regulation of the kinetic properties of transcriptional bursting

Cell-to-cell heterogeneity in gene expression can even be observed in the same type of cells, present in a similar environment. Transcriptional bursting is thought to be one of contributing factors to the heterogeneity, but it remains elusive how the kinetic properties of transcriptional bursting (e.g. burst size, burst frequency, and noise induced by transcriptional bursting) are regulated in mammalian cells. To unbiasedly identify genes regulating the kinetic properties of transcriptional bursting, we performed large-scale CRISPR/-Cas9 based screening and functional analysis, and found that Akt/MAPK signaling pathways are involved in the regulation of the kinetic properties of transcriptional bursting. Genome-wide CRISPR library screening was performed in Nanog, Dnmt3l, Trim28 knockin cell lines, in which GFP and iRFP reporter separately inserted into the two alleles. There are two technical replication in FACS-sorted sampels.