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Genome-scale mapping of DNA damage suppressors through phenotypic CRISPR-Cas9 screens.

Molecular cell (2023-07-22)
Yichao Zhao, Daniel Tabet, Diana Rubio Contreras, Linjiang Lao, Arne Nedergaard Kousholt, Jochen Weile, Henrique Melo, Lisa Hoeg, Sumin Feng, Atina G Coté, Zhen-Yuan Lin, Dheva Setiaputra, Jos Jonkers, Anne-Claude Gingras, Fernando Gómez Herreros, Frederick P Roth, Daniel Durocher
RÉSUMÉ

To maintain genome integrity, cells must accurately duplicate their genome and repair DNA lesions when they occur. To uncover genes that suppress DNA damage in human cells, we undertook flow-cytometry-based CRISPR-Cas9 screens that monitored DNA damage. We identified 160 genes whose mutation caused spontaneous DNA damage, a list enriched in essential genes, highlighting the importance of genomic integrity for cellular fitness. We also identified 227 genes whose mutation caused DNA damage in replication-perturbed cells. Among the genes characterized, we discovered that deoxyribose-phosphate aldolase DERA suppresses DNA damage caused by cytarabine (Ara-C) and that GNB1L, a gene implicated in 22q11.2 syndrome, promotes biogenesis of ATR and related phosphatidylinositol 3-kinase-related kinases (PIKKs). These results implicate defective PIKK biogenesis as a cause of some phenotypes associated with 22q11.2 syndrome. The phenotypic mapping of genes that suppress DNA damage therefore provides a rich resource to probe the cellular pathways that influence genome maintenance.

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Roche
Cocktail d'inhibiteurs de protéases sans EDTA Mini cOmplete, Protease Inhibitor Cocktail Tablets provided in a glass vial, Tablets provided in a glass vial
Sigma-Aldrich
Cocktail d'inhibiteurs de protéases, for use with mammalian cell and tissue extracts, DMSO solution
Sigma-Aldrich
Anticorps anti-phospho histone H2A.X (Ser139), clone JBW301, avec Alexa Fluor 647, clone JBW301, 0.5 mg/mL, from mouse