|Publication Type||Journal Article|
|Year of Publication||2020|
|Authors||Enache, OM, Rendo, V, Abdusamad, M, Lam, D, Davison, D, Pal, S, Currimjee, N, Hess, J, Pantel, S, Nag, A, Thorner, AR, Doench, JG, Vazquez, F, Beroukhim, R, Golub, TR, Ben-David, U|
|Date Published||2020 May 18|
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.
|Alternate Journal||Nat. Genet.|
|Grant List||Investigator grant / / Howard Hughes Medical Institute (HHMI) /|