A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens

[摘要] Mapping genetic interactions is essential for determining gene function and defining novel biological pathways. We report a simple to use CRISPR interference (CRISPRi) based platform, compatible with Fluorescence Activated Cell Sorting (FACS)-based reporter screens, to query epistatic relationships at scale. This is enabled by a flexible dual-sgRNA library design that allows for the simultaneous delivery and selection of a fixed sgRNA and a second randomized guide, comprised of a genome-wide library, with a single transduction. We use this approach to identify epistatic relationships for a defined biological pathway, showing both increased sensitivity and specificity than traditional growth screening approaches.

[发布日期] 2023-10-19

[关键词] CRISPR interference;Genetic modifier;Epistasis;Genome-wide screen;ER membrane protein complex;Tail-anchored proteins