Genome-wide RNAi screen for human 60S subunit biogenesis factors: Genome-wide RNAi screen identifies novel players in human 60S subunit biogenesis including key enzymes of polyamine metabolism

While great progress has been made in deciphering mechanisms governing ribosome biogenesis in eukaryotes, an inventory of factors that support ribosome synthesis in human cells is still missing, in particular regarding the maturation of the large 60S subunit. Here, we performed a genome-wide RNAi screen using an imaging-based, single cell assay to unravel the cellular machinery promoting 60S subunit assembly in human cells.

Two genome wide libraries from different vendors (Qiagen and Ambion) were screened in HeLa cells expressing RPL29-GFP under a tetracycline inducible promoter. Screening plates were imaged by automated microscopy (9 sites per well). Images were subjected to an automated pipeline for illimitation correction, segmentation and feature extraction.