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. 2019 Nov 6;9:16104. doi: 10.1038/s41598-019-52616-5

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© The Author(s) 2019

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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Modified tracrRNAs improve Cas9 RNP activity at other TAR crRNAs sites. The tracrRNA-UM, tracrRNA-6 and tracrRNA-19 were annealed with a TAR3,4 and 5 crRNAs and transfected into pMoHIV-C6 cells.(A) The levels of GFP expression was assessed by FACS, and (B) indel formation measured by a drop-off assay. A transfection without dgRNA (control) was included as negative control. The errors bars represent standard error of the mean (SEM) of samples treated in duplicate, and the experiment was repeated twice.*p < 0.05,**p < 0.005, ***p < 0.001 were obtained by one-way ANOVA and Dunnett’s test.