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. 2020 Jun 19;10:9957. doi: 10.1038/s41598-020-66780-6

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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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Verification of founder Il2rg-mutant rabbits using the CRISPR/Cas9 system. (a) Schematic representation of rabbit Il2rg. Magnified views illustrate the binding sites for CRISPR. (b) Co-injection of gRNA targeting rabbit Il2rg with Cas9 mRNA into rabbit male pronuclei embryos resulted mutations at the target sites. (c) PCR analysis of 30 offspring obtained through CRISPR/Cas9 injection of JW oocytes. Fourteen mutants (#2, 3, 4, 6, 8, 12, 16, 20, D1, D2, D3, D5, D8, and D9) were identified (asterisk). The product size of the wild-type JW Rabbit allele (arrow) is of 249 bp. (d) Sequencing analysis of CRISPR/Cas9-induced mutations in the target region in JW rabbits. Multiple deletions and insertions are depicted by red dashes and letters, respectively, and are aligned along the WT sequences displayed on the top line.