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. 2018 Dec 21;9:5416. doi: 10.1038/s41467-018-07901-8

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© The Author(s) 2018

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 — Comparison of CRISPRa and ORF technology. a Comparison of log2-fold change values for Calabrese Set A and Set B. Each dot represents the average of the 3 sgRNAs per gene. Top scoring genes are annotated. b Distribution of the p-values for the top 50 genes in each set of Calabrese, as well as both sets combined, for selumetinib resistance screens in MelJuSo cells. P-values were calculated using a hypergeometric distribution equivalent to a one-sided Fisher’s exact test. c Comparison of log2-fold change for MEK inhibition screens in MelJuSo cells, for an ORF library screened with trametinib and the Calabrese library screened with selumetinib. Dotted lines indicate the top 100 genes in each screen. d Comparison of the average log2-fold change values for all sgRNAs targeting a gene in the secondary pool screened with either selumetinib or trametinib. Sets of non-targeting control sgRNAs are colored in gray; dotted lines represent a 5% FDR cutoff. e Validation rate of genes in the secondary screen grouped by their p-value in the primary screen (combined sets; see (b)) for selumetinib resistance in MelJuSo cells. Number of genes per category is indicated. f Validation rate for genes grouped by the primary screen in which they scored. Dotted line indicates FDR <5%. The fraction of genes pass this cutoff is reported, as well as the total number of genes in each category. The box represents the 25th, 50th, and 75th percentiles; whiskers show 10th and 90th percentiles. For the category with 12 genes, the individual values are plotted and the mean indicated. g Secondary screen results for sgRNAs targeting BRAF. Black points are sgRNAs present in both the primary (Calabrese) and secondary pools; red points are sgRNAs that were only included in the secondary pool. h Area under the curve calculated for STOP genes in CRISPRa and held-out ORF screens. For each plot, a different ORF screen cell line (HMEC, IMR90, or HPNE) was held out, and the other two, indicated at the top of each graph, were used to generate a list of STOP genes