Genome_scale_identification_of_cellular_pathways_required_for_cell_surface_recognition. Genome_scale_identification_of_cellular_pathways_required_for_cell_surface_recognition

Interactions mediated by cell surface receptors initiate important instructive signaling cues but can be difficult to detect in biochemical assays because they are often highly transient and membrane-embedded receptors are difficult to solubilize in their native conformation. Here, we address these biochemical challenges by using a genome-scale cell-based genetic screening approach using CRISPR gene knockout technology to identify cellular pathways required for specific cell surface recognition events. Using high-affinity monoclonal antibodies and low-affinity ligands, we determined the necessary screening parameters including the importance of establishing binding contributions from the glycocalyx that permitted the unequivocal identification of genes encoding directly interacting membrane-embedded receptors with high statistical confidence. Importantly, we show that this genome-wide screening approach additionally identified receptor-specific pathways that are required for functional display of receptors on the cell surface which included chaperones, enzymes that add posttranslational modifications, trafficking proteins, and transcription factors. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/