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. 2020 Jul 10;10:11415. doi: 10.1038/s41598-020-68040-z

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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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CRISPR-Cas9-mediated disruption of zebrafish klf9. (A) Schematic of the klf9 locus and encoded protein showing location of the gRNA target in exon 1, which introduces a premature stop codon upstream of the DNA binding domain. (B) Sequence of klf9 exon 1, with 5′ untranslated region in small case and coding sequence in upper case. The location of the 2 base pair deletion generated by CRISPR is shown in gray font; the gRNA target sequence is underlined; and the premature stop codon introduced by frameshift is noted with an asterisk. (C) Effect of chronic cortisol treatment and the frameshift mutation on relative levels of klf9 mRNA and pre-mRNA, measured by qRT-PCR. The results show the averages and SEM of three biological replicates; significance values were obtained by ANOVA.