Genome-scale chemogenomic CRISPR screens in human cells using the TKOv3 library
Open accessAbstract
CRISPR-based genetic screens revolutionized our ability to genetically probe cell biology. We present a protocol to conduct genome-scale chemogenomic dropout CRISPR screens in the human RPE1-hTERT p53−/− cell line. We use the TKOv3 library, which contains 70,948 sgRNAs targeting 18,053 genes. Here, we describe how to set up the screen, the reagents required, and how to sequence and analyze the results. This protocol can be customized for other libraries, cell lines, and sequencing instruments. For complete details on the use and execution of this protocol, please refer to Olivieri et al. (2020).
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From Genome-Wide CRISPR Screens Identify Multiple Synthetic Lethal Targets That Enhance KRASG12C Inhibitor Efficacy (2023) · cites this paper as [19]
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References 9 · Cited by 38
- Authors
- Olivieri, M., & Durocher, D.
- Affiliations
- Mount Sinai Hospital (CA)
- Corresponding author
- Durocher, Daniel
- Venue
- STAR Protocols
- Publisher
- Elsevier BV
- Published
- 2021-03
- Volume
- 2
- Issue
- 1
- Pages
- 100321
- ISSN
- 2666-1667 (ISSN-L)
- PMC
- PMC7868615
- OpenAlex
- W3127207441
- Cited by
- 38
- References
- 9
- Citations by year
- 2026: 12 · 2025: 9 · 2024: 5 · 2023: 6 · 2022: 5 · 2021: 1
- FWCI
- 1.33
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- STAR Protocols (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Canadian Cancer Society
- Ontario Genomics
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chromatin Remodeling and Cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- CRISPR screen, genome-wide screen, human cells, screen analysis
- MeSH terms
- Cell Line, Humans, Gene Library, Genome, Human, CRISPR-Cas Systems
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