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

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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...

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Detalles Bibliográficos
Autores principales: Guna, Alina, Page, Katharine R., Replogle, Joseph M., Esantsi, Theodore K., Wang, Maxine L., Weissman, Jonathan S., Voorhees, Rebecca M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9882262/
https://www.ncbi.nlm.nih.gov/pubmed/36711738
http://dx.doi.org/10.1101/2023.01.22.525086
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author Guna, Alina
Page, Katharine R.
Replogle, Joseph M.
Esantsi, Theodore K.
Wang, Maxine L.
Weissman, Jonathan S.
Voorhees, Rebecca M.
author_facet Guna, Alina
Page, Katharine R.
Replogle, Joseph M.
Esantsi, Theodore K.
Wang, Maxine L.
Weissman, Jonathan S.
Voorhees, Rebecca M.
author_sort Guna, Alina
collection PubMed
description 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.
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spelling pubmed-98822622023-01-28 A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens Guna, Alina Page, Katharine R. Replogle, Joseph M. Esantsi, Theodore K. Wang, Maxine L. Weissman, Jonathan S. Voorhees, Rebecca M. bioRxiv Article 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. Cold Spring Harbor Laboratory 2023-08-14 /pmc/articles/PMC9882262/ /pubmed/36711738 http://dx.doi.org/10.1101/2023.01.22.525086 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
Guna, Alina
Page, Katharine R.
Replogle, Joseph M.
Esantsi, Theodore K.
Wang, Maxine L.
Weissman, Jonathan S.
Voorhees, Rebecca M.
A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens
title A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens
title_full A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens
title_fullStr A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens
title_full_unstemmed A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens
title_short A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens
title_sort dual sgrna library design to probe genetic modifiers using genome-wide crispri screens
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9882262/
https://www.ncbi.nlm.nih.gov/pubmed/36711738
http://dx.doi.org/10.1101/2023.01.22.525086
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