Scalable CRISPR-Cas9 chemical genetic screens in non-transformed human cells

Pooled lentiviral CRISPR-Cas9 screens are utilized for assessing the differential sensitivity or resistance of many single-gene knockouts to a compound. Here, we present a scalable approach for high-throughput compound screening by utilizing a small custom library. We describe steps to perform a pro...

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Detalles Bibliográficos
Autores principales: Lin, Kevin, Chang, Ya-Chu, Marron Fernandez de Velasco, Ezequiel, Wickman, Kevin, Myers, Chad L., Bielinsky, Anja-Katrin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9483651/
https://www.ncbi.nlm.nih.gov/pubmed/36107744
http://dx.doi.org/10.1016/j.xpro.2022.101675
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author Lin, Kevin
Chang, Ya-Chu
Marron Fernandez de Velasco, Ezequiel
Wickman, Kevin
Myers, Chad L.
Bielinsky, Anja-Katrin
author_facet Lin, Kevin
Chang, Ya-Chu
Marron Fernandez de Velasco, Ezequiel
Wickman, Kevin
Myers, Chad L.
Bielinsky, Anja-Katrin
author_sort Lin, Kevin
collection PubMed
description Pooled lentiviral CRISPR-Cas9 screens are utilized for assessing the differential sensitivity or resistance of many single-gene knockouts to a compound. Here, we present a scalable approach for high-throughput compound screening by utilizing a small custom library. We describe steps to perform a proof-of-principle chemical screen in non-transformed hTERT RPE-1 TP53(−/−) cells with higher coverage and greater timepoint resolution compared to genome-wide screens. This approach can be adapted for use in various cell lines, compounds, and other focused sgRNA libraries.
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spelling pubmed-94836512022-09-20 Scalable CRISPR-Cas9 chemical genetic screens in non-transformed human cells Lin, Kevin Chang, Ya-Chu Marron Fernandez de Velasco, Ezequiel Wickman, Kevin Myers, Chad L. Bielinsky, Anja-Katrin STAR Protoc Protocol Pooled lentiviral CRISPR-Cas9 screens are utilized for assessing the differential sensitivity or resistance of many single-gene knockouts to a compound. Here, we present a scalable approach for high-throughput compound screening by utilizing a small custom library. We describe steps to perform a proof-of-principle chemical screen in non-transformed hTERT RPE-1 TP53(−/−) cells with higher coverage and greater timepoint resolution compared to genome-wide screens. This approach can be adapted for use in various cell lines, compounds, and other focused sgRNA libraries. Elsevier 2022-09-14 /pmc/articles/PMC9483651/ /pubmed/36107744 http://dx.doi.org/10.1016/j.xpro.2022.101675 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Lin, Kevin
Chang, Ya-Chu
Marron Fernandez de Velasco, Ezequiel
Wickman, Kevin
Myers, Chad L.
Bielinsky, Anja-Katrin
Scalable CRISPR-Cas9 chemical genetic screens in non-transformed human cells
title Scalable CRISPR-Cas9 chemical genetic screens in non-transformed human cells
title_full Scalable CRISPR-Cas9 chemical genetic screens in non-transformed human cells
title_fullStr Scalable CRISPR-Cas9 chemical genetic screens in non-transformed human cells
title_full_unstemmed Scalable CRISPR-Cas9 chemical genetic screens in non-transformed human cells
title_short Scalable CRISPR-Cas9 chemical genetic screens in non-transformed human cells
title_sort scalable crispr-cas9 chemical genetic screens in non-transformed human cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9483651/
https://www.ncbi.nlm.nih.gov/pubmed/36107744
http://dx.doi.org/10.1016/j.xpro.2022.101675
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