生物
DNA修复
清脆的
基因组编辑
计算生物学
Cas9
DNA
遗传学
基因
同源重组
同源定向修复
基因组
DNA损伤
DNA错配修复
核苷酸切除修复
非同源性末端接合
染色质
突变
作者
Jeffrey A. Hussmann,Jia Ling,Purnima Ravisankar,Jun Yan,Ann Cirincione,Albert Xu,Danny Simpson,Dian Yang,Anne Bothmer,Cecilia Cotta-Ramusino,Jonathan S. Weissman,Britt Adamson
出处
期刊:Cell
[Elsevier]
日期:2021-10-28
卷期号:184 (22): 5653-5669.e25
被引量:15
标识
DOI:10.1016/j.cell.2021.10.002
摘要
Cells repair DNA double-strand breaks (DSBs) through a complex set of pathways critical for maintaining genomic integrity. To systematically map these pathways, we developed a high-throughput screening approach called Repair-seq that measures the effects of thousands of genetic perturbations on mutations introduced at targeted DNA lesions. Using Repair-seq, we profiled DSB repair products induced by two programmable nucleases (Cas9 and Cas12a) in the presence or absence of oligonucleotides for homology-directed repair (HDR) after knockdown of 476 genes involved in DSB repair or associated processes. The resulting data enabled principled, data-driven inference of DSB end joining and HDR pathways. Systematic interrogation of this data uncovered unexpected relationships among DSB repair genes and demonstrated that repair outcomes with superficially similar sequence architectures can have markedly different genetic dependencies. This work provides a foundation for mapping DNA repair pathways and for optimizing genome editing across diverse modalities.
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