Protocol of CRISPR-Cas9 knockout screens for identifying ferroptosis regulators
Summary: 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 fer...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723007293 |
| _version_ | 1797405357594640384 |
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| author | Xin Yang Shoufu Duan Zhiming Li Zhe Wang Ning Kon Zhiguo Zhang Wei Gu |
| author_facet | Xin Yang Shoufu Duan Zhiming Li Zhe Wang Ning Kon Zhiguo Zhang Wei Gu |
| author_sort | Xin Yang |
| collection | DOAJ |
| description | Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-09T03:08:48Z |
| format | Article |
| id | doaj.art-2f89bfcbcf80473abc0f63b283e474a0 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-09T03:08:48Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-2f89bfcbcf80473abc0f63b283e474a02023-12-04T05:24:07ZengElsevierSTAR Protocols2666-16672023-12-0144102762Protocol of CRISPR-Cas9 knockout screens for identifying ferroptosis regulatorsXin Yang0Shoufu Duan1Zhiming Li2Zhe Wang3Ning Kon4Zhiguo Zhang5Wei Gu6Institute for Cancer Genetics, and Herbert Irving Comprehensive Cancer Center, Vagelos College of Physicians & Surgeons, Columbia University, New York, NY, USAInstitute for Cancer Genetics, and Herbert Irving Comprehensive Cancer Center, Vagelos College of Physicians & Surgeons, Columbia University, New York, NY, USAInstitute for Cancer Genetics, and Herbert Irving Comprehensive Cancer Center, Vagelos College of Physicians & Surgeons, Columbia University, New York, NY, USAInstitute for Cancer Genetics, and Herbert Irving Comprehensive Cancer Center, Vagelos College of Physicians & Surgeons, Columbia University, New York, NY, USAInstitute for Cancer Genetics, and Herbert Irving Comprehensive Cancer Center, Vagelos College of Physicians & Surgeons, Columbia University, New York, NY, USAInstitute for Cancer Genetics, and Herbert Irving Comprehensive Cancer Center, Vagelos College of Physicians & Surgeons, Columbia University, New York, NY, USA; Department of Pediatrics, and Department of Genetics and Development, Vagelos College of Physicians & Surgeons, Columbia University, New York, NY, USA; Corresponding authorInstitute for Cancer Genetics, and Herbert Irving Comprehensive Cancer Center, Vagelos College of Physicians & Surgeons, Columbia University, New York, NY, USA; Department of Pathology and Cell Biology, Vagelos College of Physicians & Surgeons, Columbia University, New York, NY, USA; Corresponding authorSummary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723007293BioinformaticsSequence analysisCell BiologyCell cultureCell-based AssaysSequencing |
| spellingShingle | Xin Yang Shoufu Duan Zhiming Li Zhe Wang Ning Kon Zhiguo Zhang Wei Gu Protocol of CRISPR-Cas9 knockout screens for identifying ferroptosis regulators STAR Protocols Bioinformatics Sequence analysis Cell Biology Cell culture Cell-based Assays Sequencing |
| title | Protocol of CRISPR-Cas9 knockout screens for identifying ferroptosis regulators |
| title_full | Protocol of CRISPR-Cas9 knockout screens for identifying ferroptosis regulators |
| title_fullStr | Protocol of CRISPR-Cas9 knockout screens for identifying ferroptosis regulators |
| title_full_unstemmed | Protocol of CRISPR-Cas9 knockout screens for identifying ferroptosis regulators |
| title_short | Protocol of CRISPR-Cas9 knockout screens for identifying ferroptosis regulators |
| title_sort | protocol of crispr cas9 knockout screens for identifying ferroptosis regulators |
| topic | Bioinformatics Sequence analysis Cell Biology Cell culture Cell-based Assays Sequencing |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723007293 |
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