Figure 1

Generation of a nop56 mutation using the CRISPR/Cas9 system. (A) Schematic describing the target site of nop56. The genomic sequences and predicted protein sequences based on the DNA sequence in the wildtype and nop56^−/− mutants are shown. (B) WISH of nop56 in the siblings and mutants at 24 and 48 hpf. The results were verified independently three times. (C) The qRT-PCR of nop56 transcript levels in the wildtype siblings and nop56^−/− mutants at 24 and 48 hpf. The qRT-PCR was performed with biological repeats and technical repeats in triplicate (n = 3, * p < 0.05). (D–H) In vitro-transcribed nop56 mRNA (400 pg) partially rescued the phenotype in nop56 mutants. A smaller head, edema in the heart, and a shorted tail extension (arrowhead) were present. (E–I) Analysis of blood cells in the Tg(LCR:EGFP) transgenic line. The arrowhead indicates red blood cells. All scale bars represent 1 mm.