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Enache, Oana M; Rendo, Veronica; Abdusamad, Mai; Lam, Daniel; Davison, Desiree; Pal, Sangita; Currimjee, Naomi; Hess, Julian; Pantel, Sasha; Nag, Anwesha; Thorner, Aaron R; Doench, John G; Vazquez, Francisca; Beroukhim, Rameen; Golub, Todd R; Ben-David, Uri
Nature genetics, 07/2020, Letnik: 52, Številka: 7Journal Article
Cas9 is commonly introduced into cell lines to enable CRISPR-Cas9-mediated genome editing. Here, we studied the genetic and transcriptional consequences of Cas9 expression itself. Gene expression profiling of 165 pairs of human cancer cell lines and their Cas9-expressing derivatives revealed upregulation of the p53 pathway upon introduction of Cas9, specifically in wild-type TP53 (TP53-WT) cell lines. This was confirmed at the messenger RNA and protein levels. Moreover, elevated levels of DNA repair were observed in Cas9-expressing cell lines. Genetic characterization of 42 cell line pairs showed that introduction of Cas9 can lead to the emergence and expansion of p53-inactivating mutations. This was confirmed by competition experiments in isogenic TP53-WT and TP53-null (TP53 ) cell lines. Lastly, Cas9 was less active in TP53-WT than in TP53-mutant cell lines, and Cas9-induced p53 pathway activation affected cellular sensitivity to both genetic and chemical perturbations. These findings may have broad implications for the proper use of CRISPR-Cas9-mediated genome editing.
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in: SICRIS
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