Knocking out nrl diminishes but does not eradicate the rod population in zebrafish.(A) The gene structure of zebrafish nrl and the CRISPR-Cas9 target site used for knocking out nrl are shown. Orange boxes, the exons encoding the MTD and bZIP domains; red arrow, the CRISPR-Cas9 target site; black arrows, the primers used for mutation detection. (B) Sequencing validation of the homozygous nrl del8 mutation (c.230_237del8). (C) The nrl mRNA levels in the nrl-KO zebrafish were measured using qPCR. The data are shown as mean with SD (n = 3). *, p < 0.05. (D) Immunostaining of retinal sections from WT and nrl-KO zebrafish from 1 mpf to 18 mpf with the anti-Rho antibody for the rod outer segments. Scale bars: 25 μm. ROS, rod outer segment; CN, cone nuclear layer; RN, rod nuclear layer; INL, inner nuclear layer; GCL: ganglion cell layer.
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