Genome-scale mapping of DNA damage suppressors through phenotypic CRISPR-Cas9 screens.
Genome-scale mapping of DNA damage suppressors through phenotypic CRISPR-Cas9 screens.
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DOI：
10.1016/j.molcel.2023.06.025
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发表时间：
2023-07
期刊：
影响因子：
16
通讯作者：
Yichao Zhao;Daniel Tabet;Diana Rubio Contreras;Linjiang Lao;A. N. Kousholt;Jochen Weile;Henrique Melo;Lisa Hoeg;Sumin Feng;Atina G. Coté;Zhen-Yuan Lin;Dheva Setiaputra;J. Jonkers;A. Gingras;Fernando Gómez Herreros;F. Roth;D. Durocher
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文献类型：
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作者：
Yichao Zhao;Daniel Tabet;Diana Rubio Contreras;Linjiang Lao;A. N. Kousholt;Jochen Weile;Henrique Melo;Lisa Hoeg;Sumin Feng;Atina G. Coté;Zhen-Yuan Lin;Dheva Setiaputra;J. Jonkers;A. Gingras;Fernando Gómez Herreros;F. Roth;D. Durocher
为了保持基因组的完整性，细胞必须准确地复制它们的基因组，并在发生DNA损伤时修复它们。为了发现人类细胞中抑制DNA损伤的基因，我们进行了基于流式细胞术的CRISPR-Cas9筛选，以监测DNA损伤。我们确定了160个突变导致自发DNA损伤的基因，这是一个富含必需基因的列表，强调了基因组完整性对细胞适应性的重要性。我们还鉴定了227个基因，其突变导致复制干扰细胞中的DNA损伤。在鉴定的基因中，我们发现脱氧核糖-磷酸醛缩酶DERA抑制阿糖胞苷（Ara-C）引起的DNA损伤，而gnb1l（一个与22q11.2综合征有关的基因）促进ATR和相关磷脂酰肌醇3激酶相关激酶（PIKKs）的生物发生。这些结果暗示有缺陷的PIKK生物发生是与22q11.2综合征相关的一些表型的原因。因此，抑制DNA损伤的基因的表型定位为探索影响基因组维持的细胞途径提供了丰富的资源。
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 thatGNB1L, 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.