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. 2019 Jul 17;10:3144. doi: 10.1038/s41467-019-10963-x

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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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Fig. 4 — Genotoxicity with AsCpf1-based multiplexed genome editing Cells were transfected with AsCpf1–3xMYC plasmid together with either mono-cistronic guides or multiplexed guide arrays targeting non-essential genes. Each multiplexed guide array contains four different mono-cistronic guides used in this study. For example, ADAM18_multi contains 4 different guides ADAM18_sg1, ADAM18_sg2, ADAM18_sg3, and ADAM18_sg4. a Relative viability of cells treated by AsCpf1 and different single guide vectors or multiplexed guide vectors. n = 8 for transfection control, n = 4 for all mono-cistronic vectors, n = 5 for all multiplexed vectors. Error bars are present as s.d. b Phosphorylated H2A.X signal of cells treated by AsCpf1 and different single guide vectors or multiplexed guide vectors. Gemcitabine: 1-hour 100 nM gemcitabine treatment prior to In-cell Western assay as a positive control. n = 4 for all mono-cistronic vectors, n = 5 for all multiplexed vectors and transfection control. Error bars are present as s.d. c Annexin-V staining signal of cells treated by AsCpf1 and different single guide vectors or multiplexed guide vectors. n = 4 for all mono-cistronic vectors, n = 5 for all multiplexed vectors and transfection control. A.U. arbitrary units. Error bars are present as s.d. Source data are provided as a Source Data file