A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens
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12864_2023_Article_9754.pdf
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Author(s)
Guna, Alina
Page, Katharine R.
Replogle, Joseph M.
Esantsi, Theodore K.
Wang, Maxine L.
Weissman, Jonathan S.
Voorhees, Rebecca M.
Date Issued
October 30, 2023
Publisher
BioMed Central
Citation
BMC Genomics. 2023 Oct 30;24(1):651
Version
Final published version
Abstract
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.
MIT Department
Whitehead Institute for Biomedical Research
Howard Hughes Medical Institute
Massachusetts Institute of Technology. Department of Biology
Koch Institute for Integrative Cancer Research at MIT
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Creative Commons Attribution
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DOI of Published Version
https://doi.org/10.1186/s12864-023-09754-y
