Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens

Overview of the cell-based screening approach and rationale for identifying extracellular receptor-ligand interactions using CRISPR/Cas9 technology.

Prepare and characterize monomeric biotinylated proteins, including quantification and oligomerization steps to generate multivalent ligand baits for screening.

Execute genome-scale CRISPR/Cas9 screening in cell populations to identify genes whose knockout eliminates or enhances binding of the biotinylated ligand.

Analyze sequencing data from genetic screens using computational methods to identify candidate receptors and annotate related signaling pathways.

Review validated hits from genome-scale screening, highlighting cell surface binding partners identified and their functional relevance.

Conclude with key takeaways on using CRISPR screens for unbiased receptor discovery and potential downstream validation strategies.