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. 2019 Apr 30;10:401. doi: 10.3389/fgene.2019.00401

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Copyright © 2019 Zhao, Liu, Xin, Shi, Bai, Sun, Zhao, Wang, Liu, Zhao, Chen and Zhang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Construction of Rag2/IL2rg^-/- gene double-knockout mice with CRISPR/Cas9 system. Schema and sequence of sgRNAs targeted on Rag2 and IL2rg locus (A). sgRNA cleavage efficiency. Genomic template was cleaved with or without Cas9 protein and sgRNA at 37°C for 1 h; products were then identified on 1.5% agarose gel. M, 1,000 bp marker; lane 1, Rag2 template; lane 2, Rag2 template + Cas9 protein; lane 3, Rag2 template + Cas9 protein + Rag2 sgRNA; lane 4, IL2rg template; lane 5, IL2rg template + Cas9 protein; and lane 6, IL2rg template + Cas9 protein + Rag2 sgRNA (B). Genotype sequence of wild-type BALB/c and mutated mice in target region (C). mRNA relative expression of Rag2 and IL2rg in wild-type BALB/c and mutated mice (D).