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Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens
Published on: June 6, 2020
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Identification of orphan ligand-receptor relationships using a cell-based CRISPRa enrichment screening platform
Dirk H Siepe1, Lukas T Henneberg1, Steven C Wilson1
1Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States.
Elife
|September 30, 2022
Summary
Researchers developed a new cell-based screening platform to discover interactions between secreted proteins and their cell surface receptors. This method successfully identified novel ligand-receptor pairs, advancing our understanding of cell communication and providing valuable data for future research.
Area of Science:
- Proteomics and Cell Signaling
- Genomics and High-Throughput Screening
Background:
- Secreted proteins (ligands) mediate cell-cell communication, but many lack identified receptors ('orphan ligands').
- Existing high-throughput methods are not optimized for discovering extracellular protein-protein interactions.
- Cell-based screening offers a promising approach for identifying novel ligand-receptor pairs.
Purpose of the Study:
- To develop and validate a proteo-genomic cell-based CRISPR activation (CRISPRa) screening platform.
- To enable rapid, parallel deorphanization of secreted ligand-receptor interactions.
- To identify novel interactions between orphan secreted proteins and cell surface receptors.
Main Methods:
- Utilized a CRISPR activation (CRISPRa) screening platform with pooled cell surface receptor sgRNA libraries.
- Employed a magnetic bead selection-based enrichment workflow for efficient screening.
- Screened 20 secreted ligands against libraries targeting single-pass (TM1) and multi-pass (TM2+) transmembrane receptors.
Main Results:
- Identified previously unknown ligand-receptor interactions in 12 out of the performed screens.
- Validated several novel interactions using surface plasmon resonance and cell binding assays.
- Discovered three novel receptor protein tyrosine phosphatase (RPTP) ligands and a chemokine-like protein binding to killer immunoglobulin-like receptors (KIRs).
Conclusions:
- The developed CRISPRa screening platform is effective for rapid ligand-receptor deorphanization.
- The study identified several novel secreted protein and cell surface receptor interactions.
- These findings provide a valuable resource for investigating human secreted and membrane proteome interactions.
