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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. 2 — Evaluation of an alternative tracrRNA. a Comparison of the sequence of the original tracrRNA and tracr-v2. b Comparison of the log2-fold-change of perfectly matched sgRNAs and single base mismatches targeting the essential gene EEF2 for the original tracrRNA and tracr-v2. CD81-targeting sgRNAs serve as the control. Mismatched sgRNAs are binned by position, and are numbered such that nt 20 is PAM-proximal. The box represents the 25th, 50th, and 75th percentiles, and whiskers show 10th and 90th percentiles. Two-tailed Welch’s t-test was used to compare the distributions within each bin and p-values are indicated.The number of sgRNAs in each comparison is shown at the bottom. c AUC values for essential, non-essential, and non-targeting gene sets with the original tracrRNA and tracr-v2 in genome-wide viability screens. Error bars represent the range of two or three biological replicates for screens with the original and modified tracrRNA, respectively