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. 2020 Nov 9;52(11):1823–1830. doi: 10.1038/s12276-020-00521-7

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© The Author(s) 2020

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. 3 — a Procedure for assessing the cellular toxicity of different types of crRNAs. Female C57BL/6J mice were injected with the hormones pregnant mare serum gonadotropin (PMSG) and human chronic gonadotropin (hCG) before mating. Zygotes were collected from the ampullae of the oviducts and treated with RNP. Forty-eight hours post-injection, morula-stage cells were counted, and genomic DNA was prepared from the viable cells. On-target and off-target loci were amplified for deep sequencing analysis. b Comparison of the survival rates of embryos post RNP transfection. Viability was calculated by the ratio of the number of viable embryos to the total number of transfected zygotes. The total number of zygotes for each experimental batch is presented as “n”. c Viability tests for multiple target genes. OM U-rich crRNA yielded more viable cells when injected into zygotes than canonical crRNA.