Genome-scale identification of cellular pathways required for cell surface recognition.

ABSTRACT: 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. By 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 m

SUBMITTER: Sharma S 

PROVIDER: S-EPMC6120632 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature