Extracellular Interaction Screening Using CRISPR Activation

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.

Name dc:type.qualificationname

Doctor of Philosophy (PhD)

Level dc:type.qualificationlevel

Doctoral

Grantor dc:publisher.institution

University of Cambridge

Year dc:date.issued

2019

Author dc:creator

• Chong, Zheng Shan

Advisor dc:contributor.advisor

• Wright, Gavin

• CRISPR
• protein-protein interactions
• high-throughput screening

Licence

• https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/ee4d27e8-2d69-4ee2-92ac-fff451fb60bc/download
• https://www.rioxx.net/licenses/all-rights-reserved/

Language dc:language

en

DOI dc:identifier.doi

https://doi.org/10.17863/CAM.39124

OAI identifier oai:identifier

oai:www.repository.cam.ac.uk:1810/291972