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Updated: Jul 3, 2025
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Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens
Published on: June 6, 2020
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Uncovering receptor-ligand interactions using a high-avidity CRISPR activation screening platform
Liping Yang1, Timothy P Sheets2, Yang Feng1
1Tumor Angiogenesis Unit, Mouse Cancer Genetics Program (MCGP), National Cancer Institute (NCI), NIH, Frederick, MD 21702, USA.
Science Advances
|February 14, 2024
Summary
A new screening platform identifies unknown protein interactions. This discovery of siglec-4 regulating T cell activation via 4-1BB interaction advances immune checkpoint inhibitor drug development.
Area of Science:
- Immunology
- Molecular Biology
- Drug Discovery
Background:
- Most approved drugs target cell surface or secreted proteins.
- Receptor deorphanization remains a challenge, with many unknown protein binding partners.
- Identifying these interactions is crucial for developing novel therapeutics.
Purpose of the Study:
- To develop an advanced screening platform for identifying extracellular receptor-ligand interactions.
- To overcome limitations in current screening methods for weak protein interactions.
- To discover novel binding partners for uncharacterized receptors.
Main Methods:
- Combined CRISPR-CAS9 gene activation (CRISPRa) with high-avidity bead-based selection.
- Incorporated serial enrichment and flow cytometry for enhanced sensitivity.
- Applied the platform to study immune checkpoint inhibitor interactions.
Main Results:
- Successfully identified a low-affinity trans-interaction between siglec-4 and the costimulatory receptor 4-1BB.
- Demonstrated that siglec-4 regulates T cell activation.
- Achieved substantially improved screening sensitivity for weak interactions.
Conclusions:
- The developed CRISPRa-based screening platform is highly efficient for identifying extracellular interactions.
- This platform can accelerate the discovery of novel receptor-ligand partners.
- The findings pave the way for developing next-generation therapeutics, including new immune checkpoint inhibitors.
