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. 2022 Jun 18;11(1):33. doi: 10.1038/s41389-022-00410-w

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — A Overview of the CRISPR knockout screens to identify genes that differentially regulate the sensitivity of wiltdype and BRCA2-knockout HeLa cells to KU55933. B Scatterplot showing the results of genome-wide CRISPR knockout screens for cellular sensitivity to the ATM inhibitor KU55933 in wildtype and BRCA2-knockout HeLa cells. Each gene targeted by the library was ranked according to the MAGeCK score indicating genes which, when inactivated, cause specific sensitivity to HeLa-BRCA2^KO cells compared to wild-type HeLa cells. Top hits chosen for validation are indicated. C Gene Ontology analysis of the top hits with a score <0.005 which cause specific KU55933 sensitivity to BRCA2-knockout compared to wild-type HeLa cells. GO_BP terms with a negative logP of 1.85 or lower are shown. D Gene Ontology analysis of the top hits with a score < 0.005 which cause specific KU55933 sensitivity to wildtype compared to BRCA2-knockout HeLa cells. GO_BP terms with a negative logP of 1.6 or lower are shown E. Table showing the ranks, and the biological functions of the hits chosen for subsequent validation. F, G Cellular survival assays showing that knockdown of RAD17, MDC1, or USP28 results in KU55933 sensitivity in HeLa-BRCA2^KO cells (F), but not in wild-type HeLa cells (G). The average of three experiments is shown. Error bars represent standard deviations, and asterisks indicate statistical significance (t-test, two-tailed, unpaired). Western blots confirming the knockdown are shown in Supplementary Fig. S4C–E.