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Nat Genet. 2018 Aug;50(8):1132-1139. doi: 10.1038/s41588-018-0174-0. Epub 2018 Jul 27.

CRISPR-Cas9 genome editing in human cells occurs via the Fanconi anemia pathway.

Author information

1
Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
2
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
3
Department of Cell and Tissue Biology, University of California, San Francisco, San Francisco, CA, USA.
4
Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA. jcorn@berkeley.edu.
5
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA. jcorn@berkeley.edu.

Abstract

CRISPR-Cas genome editing creates targeted DNA double-strand breaks (DSBs) that are processed by cellular repair pathways, including the incorporation of exogenous DNA via single-strand template repair (SSTR). To determine the genetic basis of SSTR in human cells, we developed a coupled inhibition-cutting system capable of interrogating multiple editing outcomes in the context of thousands of individual gene knockdowns. We found that human Cas9-induced SSTR requires the Fanconi anemia (FA) pathway, which is normally implicated in interstrand cross-link repair. The FA pathway does not directly impact error-prone, non-homologous end joining, but instead diverts repair toward SSTR. Furthermore, FANCD2 protein localizes to Cas9-induced DSBs, indicating a direct role in regulating genome editing. Since FA is itself a genetic disease, these data imply that patient genotype and/or transcriptome may impact the effectiveness of gene editing treatments and that treatments biased toward FA repair pathways could have therapeutic value.

PMID:
30054595
DOI:
10.1038/s41588-018-0174-0
[Indexed for MEDLINE]

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