Chemical-genetic CRISPR-Cas9 screens in human cells using a pathway-specific library

The development of CRISPR-Cas9 screening techniques coupled with chemical inhibition of specific biological processes enables high-throughput investigation into many areas of molecular biology. We present a protocol to conduct ubiquitin proteasome system-specific chemical-genetic CRISPR-Cas9 screens...

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Detalles Bibliográficos
Autores principales: Hundley, Frances V., Toczyski, David P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8339234/
https://www.ncbi.nlm.nih.gov/pubmed/34382013
http://dx.doi.org/10.1016/j.xpro.2021.100685
Descripción
Sumario:The development of CRISPR-Cas9 screening techniques coupled with chemical inhibition of specific biological processes enables high-throughput investigation into many areas of molecular biology. We present a protocol to conduct ubiquitin proteasome system-specific chemical-genetic CRISPR-Cas9 screens in the human HAP1 cell line. This protocol can be adapted for use in other cell lines, with other compounds and types of treatments, and with any other sgRNA library. For complete details on the use and execution of this protocol, please refer to Hundley et al. (2021).

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