Work

Protocol to identify receptors of secreted proteins through CRISPR-Cas9 whole-genome screening technology

Year: 2024

Type: article

Abstract: The interaction between cell surface receptors and their ligands is crucial for intercellular communication. However, current techniques for identifying direct receptor-ligand interactions remain limited. Here, we present a protocol to identify receptors of secreted proteins using a genome-scale CRISPR-Cas9 knockout genetic screening approach. We describe steps for creating a single-guide RNA (sgRNA) lentivirus library, infecting stable Cas9-MCF7 cells, staining with tagged Cholesin, and sorting non-binding cells via flow cytometry. We then detail procedures for extracting DNA, amplifying sgRNAs, and sequencing. For complete details on the use and execution of this protocol, please refer to Hu et al. 1

Source: STAR Protocols

DOI: 10.1016/j.xpro.2024.103315

Authors: Xiaoli Hu, Yiguo Wang

Institutions: Center for Life Sciences, State Key Laboratory of Membrane Biology, Tsinghua University

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,500

Funders: National Natural Science Foundation of China, Ministry of Science and Technology of the People's Republic of China