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. 2017 Aug 23;7(8):170040. doi: 10.1098/rsob.170040

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© 2017 The Authors.

Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

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Figure 2. — Generation and identification of nps-deficient zebrafish. (a) The CRISPR/Cas9 system was used to generate npsn-knockout zebrafish. The Cas9 target was chosen as exon5 of npsn. Mutants were selected using T7E1 enzyme digestion and confirmed using Sanger sequencing. In F1 founders, a mutant line (−7, +0) containing a frameshift mutation was obtained and retained for follow-up studies. (b and c) Degradation of npsn mRNA in npsn^smu5 mutant embryos. (b) WISH results for npsn mRNA indicated significant decreases in npsn^smu5 mutant embryos at 3 dpf. (c) qRT-PCR results indicating that npsn mRNA expression was downregulated to 5% in npsn^smu5 mutant embryos. (Mean ± s.e.m., n = 20 per experiment, triplicated). Statistical significance was determined by one-way ANOVA. ***p < 0.001.