Pooled extracellular receptor-ligand interaction screening using CRISPR activation

Genome Biol. 2018 Nov 26;19(1):205. doi: 10.1186/s13059-018-1581-3.

Abstract

Extracellular interactions between cell surface receptors are necessary for signaling and adhesion but identifying them remains technically challenging. We describe a cell-based genome-wide approach employing CRISPR activation to identify receptors for a defined ligand. We show receptors for high-affinity antibodies and low-affinity ligands can be unambiguously identified when used in pools or as individual binding probes. We apply this technique to identify ligands for the adhesion G-protein-coupled receptors and show that the Nogo myelin-associated inhibitory proteins are ligands for ADGRB1. This method will enable extracellular receptor-ligand identification on a genome-wide scale.

Keywords: CRISPR activation; Cell signaling; Cell surface receptors; Extracellular protein interactions; Flow cytometry; G-protein-coupled receptor; Genome-wide screening; Monoclonal antibodies.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems
  • Genomics / methods*
  • Humans
  • Ligands
  • Receptors, Cell Surface / analysis*

Substances

  • Ligands
  • Receptors, Cell Surface