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

Author:

Guna Alina,Page Katharine R.,Replogle Joseph M.,Esantsi Theodore K.,Wang Maxine L.,Weissman Jonathan S.,Voorhees Rebecca M.

Abstract

AbstractMapping 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.

Publisher

Cold Spring Harbor Laboratory

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