Fig. 4

Generation and survival analysis of dut “crispant” zebrafish. (A) Schematic view of the zebrafish dut locus, dut transcript and the domain composition of the Dut protein. The targeting region of the dut sgRNA is highlighted. (In the genomic scheme red boxes denote exons; in the transcript view, light and dark green boxes highlight alternating exons, whereas in the schematic Dut view the yellow box denotes the dUTPase domain.) (B) The Sanger sequenogram of the target region using dut “crispant” genomic DNA from 12 embryos as a template. (C) Disruption of zygotic dut function results in embryonic lethality. Injection of Cas9 ribonucleoproteins (RNPs) targeting dut had severe effects on the estimated probability of survival of the injected embryos in two separate experiments (Group 1 and Group 2) as shown by a Kaplan–Meier survival plot. The log‐rank test was used to calculate the p‐value.