Extensible combinatorial CRISPR screening in mammalian cells

Abstract

Summary: The CRISPR-Cas system coupled with Combinatorial Genetics En Masse (CombiGEM) enables systematic analysis of high-order genetic perturbations that are important for understanding biological processes and discovering therapeutic target combinations. Here, we present detailed steps and technical considerations for building multiplexed guide RNA libraries and carrying out a combinatorial CRISPR screen in mammalian cells. We also present an analytical pipeline, CombiPIPE, for mapping two- and three-way genetic interactions.For complete details on the use and execution of this protocol, please refer to Zhou et al. (2020).

Keywords

• High Throughput Screening
• CRISPR