Genome-wide reporter screen for transcriptional regulators of ribosome biogenesis genes and ribosomal protein genes

ABSTRACT: A genome-wide CRISPR/Cas9 knockout reporter screen was performed to identify transcriptional regulators of ribosomal biogenesis genes and ribosomal protein genes. The murine GeCKO v2 library provided by the Zhang lab was amplified and positive control sgRNAs targeting the reporters were spiked-in. NIH/3T3 cells (murine fibroblasts) harboring the "ribosome biogenesis" reporter Fbl-GFP (Fibrillarin promoter-driven enhanced GFP fused to the PEST domain of the murine ornithine decarboxylase) and the "ribosomal protein" reporter Rpl18-RFP (Rpl18 promoter-driven TurboRFP fused to the PEST domain of the murine ornithine decarboxylase) were transduced with the aforementioned library, selected and FACS-sorted six days after infection. The sorted conditions were i) low GFP, but middle to strong RFP expression (RiBi down); ii) low RFP, but middle to strong GFP expression (RP down); iii) low GFP and low RFP expression (Both down); iv) strong GFP and strong RFP expression (Both up). The same amount of cells that were used for sorting, were harvested as the "unsorted" condition to which all sorted samples were compared to. The plasmid library was also sequenced.

ORGANISM(S): synthetic construct

PROVIDER: GSE207900 | GEO | 2022/07/13

REPOSITORIES: GEO