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Protocol of CRISPR-Cas9 knockout screens for identifying ferroptosis regulators.

ABSTRACT: Ferroptosis, an iron-dependent programmed cell death triggered by excessive lipid peroxidation, has shown promising therapeutic potentials in human diseases. Here, we describe a protocol of a CRISPR-Cas9 loss-of-function screen to identify regulators in response to different inducers of ferroptosis. We emphasize the steps of library amplification, drug treatment, high-throughput sequencing preparation, and bioinformatics analysis using model-based analysis of genome-wide CRISPR-Cas9 knockout (MAGeCK). We also present a method to discover the regulators of ferroptosis and verify the potential targets efficiently. For complete details on use and execution of this protocol, please refer to Yang et al. (2023).¹.

SUBMITTER: Yang X

PROVIDER: S-EPMC10730373 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature

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Protocol of CRISPR-Cas9 knockout screens for identifying ferroptosis regulators.

Yang Xin X Duan Shoufu S Li Zhiming Z Wang Zhe Z Kon Ning N Zhang Zhiguo Z Gu Wei W

STAR protocols 20231203 4

Ferroptosis, an iron-dependent programmed cell death triggered by excessive lipid peroxidation, has shown promising therapeutic potentials in human diseases. Here, we describe a protocol of a CRISPR-Cas9 loss-of-function screen to identify regulators in response to different inducers of ferroptosis. We emphasize the steps of library amplification, drug treatment, high-throughput sequencing preparation, and bioinformatics analysis using model-based analysis of genome-wide CRISPR-Cas9 knockout (MA ...[more]

PMID: 38048220

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