Details

Autor(en) / Beteiligte

• Hussmann, Jeffrey A.
• Ling, Jia
• Ravisankar, Purnima
• Yan, Jun
• Cirincione, Ann
• Xu, Albert
• Simpson, Danny
• Yang, Dian
• Bothmer, Anne
• Cotta-Ramusino, Cecilia
• Weissman, Jonathan S.
• Adamson, Britt

Titel

Mapping the genetic landscape of DNA double-strand break repair

Ist Teil von

• Cell, 2021-10, Vol.184 (22), p.5653-5669.e25

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2021

Quelle

Elsevier ScienceDirect Journals

Beschreibungen/Notizen

• 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. [Display omitted] •Repair-seq maps the genetic dependencies of DNA repair outcomes•High-resolution signatures of gene function identify unexpected gene relationships•DSB-induced mutations with similar sequences can result from distinct mechanisms•Repair-seq can be adapted to study a broad range of genome editing tools Measuring the effects of many genetic perturbations on the spectrum of mutations produced at targeted DNA breaks allows systematic mapping of DNA repair pathways.