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
Descripción
Sumario: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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