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

Kevin Lin; Ya Chu Chang; Ezequiel Marron Fernandez de Velasco; Kevin Wickman; Chad L. Myers; Anja K Bielinsky

doi:10.1016/j.xpro.2022.101675

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

Kevin Lin, Ya Chu Chang, Ezequiel Marron Fernandez de Velasco, Kevin Wickman, Chad L. Myers, Anja K Bielinsky

Pharmacology

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

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.

Original language	English (US)
Article number	101675
Journal	STAR Protocols
Volume	3
Issue number	4
DOIs	https://doi.org/10.1016/j.xpro.2022.101675
State	Published - Dec 16 2022

Bibliographical note

Publisher Copyright:
© 2022 The Author(s)

Keywords

CRISPR
Genomics
High Throughput Screening
Molecular Biology
Sequencing

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abstract = "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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AU - Chang, Ya Chu

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AU - Wickman, Kevin

AU - Myers, Chad L.

AU - Bielinsky, Anja K

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KW - Molecular Biology

KW - Sequencing

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

Abstract

Bibliographical note

Keywords

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