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

ABSTRACT: 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.

SUBMITTER: Guna A

PROVIDER: S-EPMC9882262 | biostudies-literature | 2023 Aug

REPOSITORIES: biostudies-literature

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

Guna Alina A Page Katharine R KR Replogle Joseph M JM Esantsi Theodore K TK Wang Maxine L ML Weissman Jonathan S JS Voorhees Rebecca M RM

bioRxiv : the preprint server for biology 20230814

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 lib ...[more]

PMID: 36711738

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2022-06-04 | GSE205310 | GEO

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